tag:blogger.com,1999:blog-85008081378766376142024-02-20T08:33:34.914-08:00Summer Research with the College of PharmacyLauren Tolathttp://www.blogger.com/profile/05812297914953765766noreply@blogger.comBlogger12125tag:blogger.com,1999:blog-8500808137876637614.post-39245719260292095602012-08-10T08:54:00.000-07:002012-08-10T08:55:25.355-07:00Week 12-Conclusion to my Summer Blogging!This week was supposed to be the last official week of summer research (as per the UH academic calendar); however, I am so pleased with my experience in Dr. McConnell's lab that we both agreed to continue my position in his lab throughout the fall and spring semesters (at reduced hours and reduced days, of course). This is important to me because I want to apply and continue what I have learned during this internship to my own independent research project (and perhaps honors thesis) in the future. I feel as if the skills and technique I have picked up during this experience are extremely invaluable to any lab setting I may encounter, whether it be a course-required lab or a lab used in the institutional setting. One thing I really enjoyed learning was the actual practice of asceptic technique in a laminar flow hood, something that my supplemental studies indicated was extremely important in the pharmacy setting. My graduate student mentor will still be working towards her Ph.D. and the rest of the research team will still be continuing their experiments by the time the fall semester kicks up, so I will be stopping by to help them with the same things I have learned this summer.<br />
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For the next few work-days, I will be helping one of the members of the research team with running cAMP and PKA (with PKI) assays. This will be used to determine cAMP (cyclic AMP) levels in our tissues of interest.<br />
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My graduate student mentor has also been showing me her technique for calcium imaging to measure the intercellular calcium release in our cells of interest.<br />
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The lab has been quite hectic because we are within weeks of revising and submitting over six papers for publication. Despite this though, it is the unity that comes about in times like this that never ceases to make me proud that I am a part of such a hard-working lab.<br />
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Thank you to my Tier One Scholarship, the University of Houston, and Dr. McConnell for this amazing opportunity!<br />
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<table align="center" cellpadding="0" cellspacing="0" class="tr-caption-container" style="margin-left: auto; margin-right: auto; text-align: center;"><tbody>
<tr><td style="text-align: center;"><a href="https://sphotos-b.xx.fbcdn.net/hphotos-snc7/s720x720/600112_353828284692020_141293957_n.jpg" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="400" src="https://sphotos-b.xx.fbcdn.net/hphotos-snc7/s720x720/600112_353828284692020_141293957_n.jpg" width="290" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Our Lab's Updated Roster</td></tr>
</tbody></table>Lauren Tolathttp://www.blogger.com/profile/05812297914953765766noreply@blogger.com9tag:blogger.com,1999:blog-8500808137876637614.post-13781521572369493802012-07-27T12:46:00.005-07:002012-07-27T13:24:38.022-07:00Week 10 & 11-The Research ContinuesThis week has been the busiest by far for me. Since I'll be out of town next week, I have ensured that all of my duties have been performed in such a way to last an extra week. Western blots, cardiomyocyte isolations, buffer preparation, lipid lesion analysis, and surgeries on our specimen are still running rampant in the lab. <br />
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After watching my <a href="http://vimeo.com/46012623">DNA Gel Electroporesis video</a>, Dr. McConnell, my PI, suggested that I create a library of lab videos to show incoming students as well as students who are undergoing their rotations within the department the proper technique for basic lab functions. <br />
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<iframe allowfullscreen="" frameborder="0" height="375" mozallowfullscreen="" src="http://player.vimeo.com/video/46405084" webkitallowfullscreen="" width="500"></iframe> <br />
<a href="http://vimeo.com/46405084">Storing Western Blot Membranes-Lauren Tolat</a> from <a href="http://vimeo.com/user11736417">Lauren Tolat</a> on <a href="http://vimeo.com/">Vimeo</a>.<br />
<iframe allowfullscreen="" frameborder="0" height="375" mozallowfullscreen="" src="http://player.vimeo.com/video/46409568" webkitallowfullscreen="" width="500"></iframe> <br />
<a href="http://vimeo.com/46409568">How to: Standardize and Use the pH Meter - Lauren Tolat</a> from <a href="http://vimeo.com/user11736417">Lauren Tolat</a> on <a href="http://vimeo.com/">Vimeo</a>.<br />
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The first video demonstrates the technique (and some tips) involved in storing Western Blot membranes. A common way to store the membranes is by transferring them to a polyethylene bag and keeping them saturated in TBST. The final product is stored in the refrigerator (2 to 8 degrees Celsius).<br />
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The second video is a technique used in general lab settings-standardizing and using the pH meter. When preparing a high volume of buffer solutions that need specific pH and temperature adjustments, the pH meter is a lifesaver. However, many lab students are unaware of the lengthy procedure involved in standardizing and cautiously using the meter for their own measurements. <br />
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Overall, I am beyond ecstatic about ALL of the opportunities that I have been able to experience during my undergraduate research and I feel incredibly blessed to be here at the University of Houston.Lauren Tolathttp://www.blogger.com/profile/05812297914953765766noreply@blogger.com0tag:blogger.com,1999:blog-8500808137876637614.post-35186230068719414282012-07-20T10:39:00.000-07:002012-07-20T12:51:32.038-07:00Week 9-DNA Gel ElectrophoresisThis week has been super busy! Among cardiomyocyte isolations, western blots, protein assays, operations on our live specimen, and a potluck party thrown in honor of my graduate student mentor's successful research proposal, I had the opportunity to perform my very own DNA Gel Electrophoresis. Electrophoresis is a method used in this lab for genotyping among our six different experimental groups. Specifically, it is used to determine whether our gene of interest is available in a particular sample after PCR has been performed. It involves four steps: 1) preparing the gel; 2) loading the gel; 3) running the gel; and, 4) visualizing. With the lovely help of my multi-talented research team, I created a video of me performing these steps.<br />
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<iframe allowfullscreen="" frameborder="0" height="375" mozallowfullscreen="" src="http://player.vimeo.com/video/46012623" webkitallowfullscreen="" width="500"></iframe><br />
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgVQX9F5B40F10L8AniYHcaN2BqUL0Z_vPlCniBptz3oaohWiw3tFZMCc7fgNwdmlpjcyJFcRHpq7RYpxl5wP6wePhKXd7_R3t-DSKMAphqCqCWYqMkbrZDjCV9y0rzGKDn65auRtbOdHk/s1600/P7170022.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgVQX9F5B40F10L8AniYHcaN2BqUL0Z_vPlCniBptz3oaohWiw3tFZMCc7fgNwdmlpjcyJFcRHpq7RYpxl5wP6wePhKXd7_R3t-DSKMAphqCqCWYqMkbrZDjCV9y0rzGKDn65auRtbOdHk/s200/P7170022.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">An Assortment of Food-Potluck Party</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjQ0t1r8gPiOVYMzZjY_1AiNKngNMPsrTLDns5gleXrfu-pqKqKy6WhhnXFbZ0lXtyz8DgBO7q21nGTrBWO6BVlA1KQ8dwYA7YqOHShvL38sHFhgiQKDwG_1u0pUwCyl18h7JBqivFyzlM/s1600/P7170025.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjQ0t1r8gPiOVYMzZjY_1AiNKngNMPsrTLDns5gleXrfu-pqKqKy6WhhnXFbZ0lXtyz8DgBO7q21nGTrBWO6BVlA1KQ8dwYA7YqOHShvL38sHFhgiQKDwG_1u0pUwCyl18h7JBqivFyzlM/s200/P7170025.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">The Team-Potluck Party</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhQPH8gss42RIsccmt9U46eqrHJ6vsFVlFDLNdNKlros-MRkM7gXARCvW-GDqBNzk_M2c7wt_d8LhIUA4e7Ecm4pihxoU2mttRcoR1pGqK_fT3vtBh_mles2uS7YddXJ2LA9j-7jlEFQhw/s1600/P7170026.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhQPH8gss42RIsccmt9U46eqrHJ6vsFVlFDLNdNKlros-MRkM7gXARCvW-GDqBNzk_M2c7wt_d8LhIUA4e7Ecm4pihxoU2mttRcoR1pGqK_fT3vtBh_mles2uS7YddXJ2LA9j-7jlEFQhw/s200/P7170026.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">The Team-Potluck Party</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgUJrQL5B_61cawNXwohcYXdyjromguMPdwZJoniSG-_TJ4fiutxb3UhubMq0yNAhJ5-ad4_ELO5cVhNqelB3LF7eFSL6ulEDMyJ2SSVWQ-ch_GRWX-jGLHRvkyq57X-kXN3FlD56Oq_28/s1600/P7170027.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgUJrQL5B_61cawNXwohcYXdyjromguMPdwZJoniSG-_TJ4fiutxb3UhubMq0yNAhJ5-ad4_ELO5cVhNqelB3LF7eFSL6ulEDMyJ2SSVWQ-ch_GRWX-jGLHRvkyq57X-kXN3FlD56Oq_28/s200/P7170027.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Cori and Fan</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjNr2p9nbyWK63Dkev053GH3rJ3JWq7dXHu69PfxIT2mfIwRiGcWhmSKcAkdmlNCW3KBoLDHwkNiqgd4CItkX9ErmGaac3q5ZsM6UCxSxY4w2UV-AHgEQMIEXD37Ejk89XpKyNUUqlCj-Q/s1600/P7170028.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjNr2p9nbyWK63Dkev053GH3rJ3JWq7dXHu69PfxIT2mfIwRiGcWhmSKcAkdmlNCW3KBoLDHwkNiqgd4CItkX9ErmGaac3q5ZsM6UCxSxY4w2UV-AHgEQMIEXD37Ejk89XpKyNUUqlCj-Q/s200/P7170028.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Western Blots</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEh9OfIz8XQt3grnpiRCLu1qZ11UF2x_Ip_wSYb0zXjGeV6VN__85kU7ukrJVPlYx6My4JFhIKGragI5kr487Vnw1v0EZJdYZNl8JFPwLTjGxheflHMW6jtncw7aaWR2J_ow8bBaTOofH5E/s1600/P7170030.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEh9OfIz8XQt3grnpiRCLu1qZ11UF2x_Ip_wSYb0zXjGeV6VN__85kU7ukrJVPlYx6My4JFhIKGragI5kr487Vnw1v0EZJdYZNl8JFPwLTjGxheflHMW6jtncw7aaWR2J_ow8bBaTOofH5E/s200/P7170030.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">An Alternative to Milk in Western Blotting-BSA</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhJQBuTfdnyVWxjse-W-R4Jwj3Ptozr84BasTHYN5FV0CCjCrat1jCVW4QOQBGqdzd1oS9-6Bl-uNA9wZwCyy9pct8AOXcgwqD9_2QFMHCpGE2eD432UKzc1xA4y0QfDlEUZhaw3wOGHRA/s1600/P7170031.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhJQBuTfdnyVWxjse-W-R4Jwj3Ptozr84BasTHYN5FV0CCjCrat1jCVW4QOQBGqdzd1oS9-6Bl-uNA9wZwCyy9pct8AOXcgwqD9_2QFMHCpGE2eD432UKzc1xA4y0QfDlEUZhaw3wOGHRA/s200/P7170031.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Lauren's Homemade Stripping Buffer :)</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiIUqX8R1U9OckyiPxOhcU2iOxNPr5R2J92TV87cBuVGnoLX1ATeK5AP4IO6kuRfaRPDa66BwV5BwrTr3Yf-RvbYuMNq_bvKYdkOAZSlsFwBTjAjXkOYAwl8xGgEWl2ppyDsaN_3dVa49Q/s1600/P7180034.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiIUqX8R1U9OckyiPxOhcU2iOxNPr5R2J92TV87cBuVGnoLX1ATeK5AP4IO6kuRfaRPDa66BwV5BwrTr3Yf-RvbYuMNq_bvKYdkOAZSlsFwBTjAjXkOYAwl8xGgEWl2ppyDsaN_3dVa49Q/s200/P7180034.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Casket used to Mold Gel</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEh6nlXCQlxivd9YPT8dtXPApSo_Uh5rurpWAzxs9VYyb1uprpflmWF5MjZfSMXHar8hc7o9bv5dYFo4g3cMloiddGUCGZwMpCK_6IzNrmZxHIooTrrqqwkqaeusLboCKO_29QDudRRQm-s/s1600/P7180046.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEh6nlXCQlxivd9YPT8dtXPApSo_Uh5rurpWAzxs9VYyb1uprpflmWF5MjZfSMXHar8hc7o9bv5dYFo4g3cMloiddGUCGZwMpCK_6IzNrmZxHIooTrrqqwkqaeusLboCKO_29QDudRRQm-s/s200/P7180046.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Running the Gel</td></tr>
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<table align="center" cellpadding="0" cellspacing="0" class="tr-caption-container" style="margin-left: auto; margin-right: auto; text-align: center;"><tbody>
<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEihbNX7ewJB1Ac9FOpEJ9elv1cCVR7tL4A3HGugUQ6IQ2zsRgd2DPZOAM45gwBE3mPIlWJdEZKoHyT4Ooyb11ywwAqy7W-z6gaaOZujrr-fQdKK-g_PGCt4nbn_mRmrA25Q6Wvng4bbYVU/s1600/P7180048.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEihbNX7ewJB1Ac9FOpEJ9elv1cCVR7tL4A3HGugUQ6IQ2zsRgd2DPZOAM45gwBE3mPIlWJdEZKoHyT4Ooyb11ywwAqy7W-z6gaaOZujrr-fQdKK-g_PGCt4nbn_mRmrA25Q6Wvng4bbYVU/s200/P7180048.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Protein Electrophoresis-Loading Samples</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgNqohpCyXd71Ou7bWgmOpBMhGcsCvC3s2srdWyZZfph7rAxHGq_9UXyCMXihzliRl77I37TL37n9mTp7_G7dwMv_UUfWPajVUQDPk7KRrxvAHWgpL6Vv78-pVliMhnbJY3doPPPt_hkNs/s1600/P7180049.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgNqohpCyXd71Ou7bWgmOpBMhGcsCvC3s2srdWyZZfph7rAxHGq_9UXyCMXihzliRl77I37TL37n9mTp7_G7dwMv_UUfWPajVUQDPk7KRrxvAHWgpL6Vv78-pVliMhnbJY3doPPPt_hkNs/s200/P7180049.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Checking the Current</td></tr>
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<table align="center" cellpadding="0" cellspacing="0" class="tr-caption-container" style="margin-left: auto; margin-right: auto; text-align: center;"><tbody>
<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhOLn_h3hiseK1J5hLjkp6Hf3vK3CfZKM5RUVxZ5C0vtl2paoqu3QnG1tICHDaoY5LOOr2WoV1kAzUrABzf_TW8CNizn1Ul5FAH2XR_EupZYwQcJpO_5zrT-N7gTUrWQ6gGMhzmo7pK-kU/s1600/P7180051.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhOLn_h3hiseK1J5hLjkp6Hf3vK3CfZKM5RUVxZ5C0vtl2paoqu3QnG1tICHDaoY5LOOr2WoV1kAzUrABzf_TW8CNizn1Ul5FAH2XR_EupZYwQcJpO_5zrT-N7gTUrWQ6gGMhzmo7pK-kU/s200/P7180051.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;"><span style="background-color: white;">Light Cabinet-Visualizing </span></td></tr>
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<table align="center" cellpadding="0" cellspacing="0" class="tr-caption-container" style="margin-left: auto; margin-right: auto; text-align: center;"><tbody>
<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEj4HKtTUEA_ulk79FIF7THtfizwqKKRSZg_t9Fc4mESXeXM2kKAQls5faIfZmqVcuyJtMNG4jrlUaJE-vAzwY9UGedt_jSq7mbMRl3Z1fw4jyoRsk7ZJpd_Od2SN8LLiCo6sGSAXhL1T8A/s1600/P7180052.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEj4HKtTUEA_ulk79FIF7THtfizwqKKRSZg_t9Fc4mESXeXM2kKAQls5faIfZmqVcuyJtMNG4jrlUaJE-vAzwY9UGedt_jSq7mbMRl3Z1fw4jyoRsk7ZJpd_Od2SN8LLiCo6sGSAXhL1T8A/s200/P7180052.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Light Cabinet-Visualizing</td></tr>
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<table align="center" cellpadding="0" cellspacing="0" class="tr-caption-container" style="margin-left: auto; margin-right: auto; text-align: center;"><tbody>
<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEidZ8_ojvSb4RSbhSUpHCDDmCUngP5NI16NYri1G8H-e_0D8ePohdSmsz3hSXMz3xgBW8TUtpcL-QUu5Rme928vI3yNfCatmJhusdWmhG5RuoAaf8yGc00zOXPn8zpQQW1BakpcGYkgP-k/s1600/P7180053.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEidZ8_ojvSb4RSbhSUpHCDDmCUngP5NI16NYri1G8H-e_0D8ePohdSmsz3hSXMz3xgBW8TUtpcL-QUu5Rme928vI3yNfCatmJhusdWmhG5RuoAaf8yGc00zOXPn8zpQQW1BakpcGYkgP-k/s200/P7180053.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Software used in Electrophoresis</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhVmPseH6gWCNb8QUAmMQ25DDFUn9rDRIb9KGPIyEkVkXg-1pxf9r8zqK8nxl-guX_mrKsjmbYV1YYi4ylJyyPMjh8VvXrBnpN3wGCxMr19faFRb7mJU46dSUp-AItGt3J9PrXoiCFvliY/s1600/P7180056.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhVmPseH6gWCNb8QUAmMQ25DDFUn9rDRIb9KGPIyEkVkXg-1pxf9r8zqK8nxl-guX_mrKsjmbYV1YYi4ylJyyPMjh8VvXrBnpN3wGCxMr19faFRb7mJU46dSUp-AItGt3J9PrXoiCFvliY/s200/P7180056.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Visualizing the Bands</td></tr>
</tbody></table>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEh1xmyEeiH36mPurrACTCx2_0tb6l0N5xNH5PDv_LXRGf2PnAEB5hpo1QQ5T7JATa5E3Sh4zui7Qe4imWTR-8ePpulu3aWCN90ynPWBXf3EvOUF4OI-6cN4_XxF5jXgN1L6JsOqaMjJ4_o/s1600/P7180058.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEh1xmyEeiH36mPurrACTCx2_0tb6l0N5xNH5PDv_LXRGf2PnAEB5hpo1QQ5T7JATa5E3Sh4zui7Qe4imWTR-8ePpulu3aWCN90ynPWBXf3EvOUF4OI-6cN4_XxF5jXgN1L6JsOqaMjJ4_o/s200/P7180058.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Choosing a Filter to View the Results of Electrophoresis</td></tr>
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<table align="center" cellpadding="0" cellspacing="0" class="tr-caption-container" style="margin-left: auto; margin-right: auto; text-align: center;"><tbody>
<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEizV4fUJhEkEzqODh8P9nBND4stoFW9AuK2oQyoX7qs-seKzhxu6icWzoBEOvItifGCDIWM-uW8NXgr4_uJR_YS0s7stGBV-_EGNfRZb7kWew81tePiJnRkdJiGyLBk3G4jq1eNwwTEhu0/s1600/P7190066.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEizV4fUJhEkEzqODh8P9nBND4stoFW9AuK2oQyoX7qs-seKzhxu6icWzoBEOvItifGCDIWM-uW8NXgr4_uJR_YS0s7stGBV-_EGNfRZb7kWew81tePiJnRkdJiGyLBk3G4jq1eNwwTEhu0/s200/P7190066.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Transferring Membranes to Polyethylene Bags</td></tr>
</tbody></table>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgNlT3es9K4My2BOxrGa_siSf2e_Xr7_TcJ2xzEMnL3CLMDe2l78aBk3HAGMLzZwOHZMQw6_7FD4gc2e8B3TUgnKCRuR3zlMtSNyD-Fz7PV7CVC0NZB8xg0q6MIrjCAeSrPMhKu0fEFdg8/s1600/P7190067.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgNlT3es9K4My2BOxrGa_siSf2e_Xr7_TcJ2xzEMnL3CLMDe2l78aBk3HAGMLzZwOHZMQw6_7FD4gc2e8B3TUgnKCRuR3zlMtSNyD-Fz7PV7CVC0NZB8xg0q6MIrjCAeSrPMhKu0fEFdg8/s200/P7190067.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;"><span style="background-color: white;">Transferring Membranes </span></td></tr>
</tbody></table>
<table align="center" cellpadding="0" cellspacing="0" class="tr-caption-container" style="margin-left: auto; margin-right: auto; text-align: center;"><tbody>
<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEicVuTVSpwT3-rR7BSZZ3FYCitTOwU0T9sj0frMIQyf3EHoIJqMdRsFA7ICwRnxPDEZRNXDI_qHQUSqApJyc-2Gv5I53cpdncWgyhpLQu52ZMibKoPy0Ttw7_GPOk_PLMLdTn9VHt0EJ_4/s1600/P7190068.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="200" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEicVuTVSpwT3-rR7BSZZ3FYCitTOwU0T9sj0frMIQyf3EHoIJqMdRsFA7ICwRnxPDEZRNXDI_qHQUSqApJyc-2Gv5I53cpdncWgyhpLQu52ZMibKoPy0Ttw7_GPOk_PLMLdTn9VHt0EJ_4/s200/P7190068.JPG" width="150" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Storing Membranes</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjlKDirRZizx9jWO5GZn_6oU675FoyCB_0mQUtaNUT-uCudscjsLKUovIK3n585sCPpepXcBhyhtRm3loWJEMJUOi9ZlrdMggDSHTY8KmJblyaMwltTIGwx4D7lPtgc9-HSE3vyRP4VIbI/s1600/P7200071.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjlKDirRZizx9jWO5GZn_6oU675FoyCB_0mQUtaNUT-uCudscjsLKUovIK3n585sCPpepXcBhyhtRm3loWJEMJUOi9ZlrdMggDSHTY8KmJblyaMwltTIGwx4D7lPtgc9-HSE3vyRP4VIbI/s200/P7200071.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Preparing Hypotonic Buffer for Cell Culture</td></tr>
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<br />Lauren Tolathttp://www.blogger.com/profile/05812297914953765766noreply@blogger.com0tag:blogger.com,1999:blog-8500808137876637614.post-65199537479072682522012-07-13T13:05:00.002-07:002012-07-13T15:27:06.939-07:00Week 8-Solutions, Buffers, and the Joy of Scientific ReplicationCongratulations are in order for Ms. Qiying Fan, the hard-working and dedicated Ph.D. graduate student, who successfully completed her graduate research proposal this week. The entire research team is so proud of her and we couldn't have asked for a more positive response from the committee.<br />
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiGMa06HLMrAv-KorB0oD1kA9xWLu44SquUWahqQHXbAfoPR-3cfR6b7syYktjLeEwRpqofmTZxmC-wLA1JhbYQN3OMKfqtydCpF5YdjOUtIqupDWTdT9oAqIv1YPJIE1e669sg-NkpqIk/s1600/P7120014.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiGMa06HLMrAv-KorB0oD1kA9xWLu44SquUWahqQHXbAfoPR-3cfR6b7syYktjLeEwRpqofmTZxmC-wLA1JhbYQN3OMKfqtydCpF5YdjOUtIqupDWTdT9oAqIv1YPJIE1e669sg-NkpqIk/s200/P7120014.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Congrats to Fan!</td></tr>
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<span style="background-color: white;">The entire research team and our PI have been pretty busy-writing grants and conducting/planning experiments. </span><br />
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhEL1zVCZIwuoGZfpkAusoHNxhULXVvuVqoWybFmu5bjCWxcoNG77yHonuHoT61buA4b_pBgNiBwo170sFhBJ2RY4vEIssEkR0QMYl7oGXrlrao2qdzx-cJOmQIqW14YAQSGSFe7tenhl0/s1600/P7100002.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhEL1zVCZIwuoGZfpkAusoHNxhULXVvuVqoWybFmu5bjCWxcoNG77yHonuHoT61buA4b_pBgNiBwo170sFhBJ2RY4vEIssEkR0QMYl7oGXrlrao2qdzx-cJOmQIqW14YAQSGSFe7tenhl0/s200/P7100002.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Tony Ohonba-Pharmacy Student </td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEikBTkEwE8ujrdcg61j7_2_JoWgfbfZLWL2ZJEbidTNHoUBBkOw_Jgg3fYLmy6Loow3CfbGmd2bDRAswznTx6cxa6FCsbUiORyyTILQQ_F9zxGDJty2Krb5dl1njdjNg7wBQMLAzQSBnus/s1600/P7100003.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEikBTkEwE8ujrdcg61j7_2_JoWgfbfZLWL2ZJEbidTNHoUBBkOw_Jgg3fYLmy6Loow3CfbGmd2bDRAswznTx6cxa6FCsbUiORyyTILQQ_F9zxGDJty2Krb5dl1njdjNg7wBQMLAzQSBnus/s200/P7100003.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">My Graduate Student Mentor and I</td></tr>
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<span style="background-color: white;">Andrea has been planning experiments and the next steps for her project which is likely to span months. During the previous weeks, she had been refining her skills at inducing myocardial infarctions on her live specimen (mice). Now, with her technique finalized, she is officially ready to begin the actual experimental part of the project. She will be injecting certain stem cells into her specimen to test whether these cells improve cardiac function after the infarction. Her overreaching goal is to figure out if these stems cells aid in the recovery of localized damaged tissue in the heart.</span><br />
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Abeer has been continuing her experiments involving cardiomyocyte isolations. Once she obtains the cardiomyocytes (the cells that comprise cardiac muscle), she performs measurements to figure out the calcium ratio present using a specialized miscroscope called the Ionoptix. The calcium ratio will determine the degree of contractility in the cardiomyocytes.<br />
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Fan, while still recovering from a stressful week, has been continuing the development of Western blots to detect different treatments of Bapta-AM, a calcium inhibitor (used because calcium is involved in the specific pathway she is trying to identify). She has been treating VSMC (vascular smooth muscle cells) to see how specific growth factors affect VSMC intracellular signaling. Overall, this will further her understanding of how calcium, and how much with her different experimental groups, is involved in the pathway.<br />
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I had to opportunity to make my own Western Blot Stripping Buffer using substrates from the lab. A stripping buffer is used in Western blots before the blocking stage to remove primary and secondary antibodies from the PVDF membrane and allow chemiluminescent Western blots to be reprobed. This saves time and is more efficient than continuously performing gel electrophoresis and duplicate immunoblot assays. In other words, it allows the experimenter to reuse a PVDF blot to detect a different target with a different primary antibody.<br />
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<tr><td style="text-align: center;"><a href="http://www.piercenet.com/media/Restore-Western-blot-Strip-Reprobe-1.jpg" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="200" src="http://www.piercenet.com/media/Restore-Western-blot-Strip-Reprobe-1.jpg" width="99" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Effect of Stripping Buffer on Membranes</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEi0n09bwMtCTSo6a6e-EAzYbZhU2NqksTxV5P2TbOhP95TDJgYrxHGT_lGHUAZ3SoeNQf8o32wXm-dnhdF3x31Qs8UoyKkOBHmcV7b4b7gvY3ajW6bFKstwna3yRwLWaeoa_oOrAv8hsq4/s1600/P7110004.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEi0n09bwMtCTSo6a6e-EAzYbZhU2NqksTxV5P2TbOhP95TDJgYrxHGT_lGHUAZ3SoeNQf8o32wXm-dnhdF3x31Qs8UoyKkOBHmcV7b4b7gvY3ajW6bFKstwna3yRwLWaeoa_oOrAv8hsq4/s200/P7110004.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Mild Stripping Buffer-Protocol</td></tr>
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<a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEi1DXTTI8IgMg_kAV1FgVTa5-tCqPkOZwdG3ZXxQf0oWo32BrmOvxRKpHOuPVlqJrdw8qbiRO_FnB_VOY4FcvZLT9yUXf5GimJT4Gg1OD6nz_z4atOgdwX5jJQ0k7PRXVfDfuEt1ZY8qxQ/s1600/P7110006.JPG" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEi1DXTTI8IgMg_kAV1FgVTa5-tCqPkOZwdG3ZXxQf0oWo32BrmOvxRKpHOuPVlqJrdw8qbiRO_FnB_VOY4FcvZLT9yUXf5GimJT4Gg1OD6nz_z4atOgdwX5jJQ0k7PRXVfDfuEt1ZY8qxQ/s200/P7110006.JPG" width="200" /></a></div>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjXNJd5xkaW0JLLVpQjsSrWdituK1UnvbIB9sChlxqc7Gsz86oDoAsOy0sqWm_8U-1BtXaackB49q2CDay-9pJVeO4wdzVW49mcWmf-jCwmaYus8uSKlACVNX6rCzwEoYImSxQcmYgPORo/s1600/P7110005.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="240" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjXNJd5xkaW0JLLVpQjsSrWdituK1UnvbIB9sChlxqc7Gsz86oDoAsOy0sqWm_8U-1BtXaackB49q2CDay-9pJVeO4wdzVW49mcWmf-jCwmaYus8uSKlACVNX6rCzwEoYImSxQcmYgPORo/s320/P7110005.JPG" width="320" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Glycine-15 g</td></tr>
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<a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgf9t4UTWDqrr6sJu9yiKPsLkU4vr8knJWSWJ6l59Ak-sOXlpGdANe5sKSsbsKahl_9e5i8ruZP88TjQiiYb5lMG1F1TRjzjRJWWqQIsMoqbWVqAV-vrYmHbxzMwEfFhwiHwkP3JgOXVFM/s1600/P7110007.JPG" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" height="320" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgf9t4UTWDqrr6sJu9yiKPsLkU4vr8knJWSWJ6l59Ak-sOXlpGdANe5sKSsbsKahl_9e5i8ruZP88TjQiiYb5lMG1F1TRjzjRJWWqQIsMoqbWVqAV-vrYmHbxzMwEfFhwiHwkP3JgOXVFM/s320/P7110007.JPG" width="240" /></a></div>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgH5kjiZagCrqCJec_p2TDzhhPDLb9eyWt7TJYPquzRE9Xc7uS58QnfEsaCrulsTw6xIG6QYrX9g2md2ONFTJrjCzUC8uDFVhVfj2RR90OzwcfLxOvOyaU53xFhtCiahs9g_Z5mjlK8NyA/s1600/P7110008.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="240" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgH5kjiZagCrqCJec_p2TDzhhPDLb9eyWt7TJYPquzRE9Xc7uS58QnfEsaCrulsTw6xIG6QYrX9g2md2ONFTJrjCzUC8uDFVhVfj2RR90OzwcfLxOvOyaU53xFhtCiahs9g_Z5mjlK8NyA/s320/P7110008.JPG" width="320" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">SDS-1 g</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiSsEEg7AtbNqhMqwhOXl4PLJPOaW5CrKVb0UiIb5xaLhndf_zCqSMofmUDoTABJbjrrRHwan8nidLL3jgcMNRbrA51OUg4vqhUzZ9Y5vAaMj83RI0wjuBVtolROCJEc1e8PnbxllOMD7Y/s1600/P7110009.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="240" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiSsEEg7AtbNqhMqwhOXl4PLJPOaW5CrKVb0UiIb5xaLhndf_zCqSMofmUDoTABJbjrrRHwan8nidLL3jgcMNRbrA51OUg4vqhUzZ9Y5vAaMj83RI0wjuBVtolROCJEc1e8PnbxllOMD7Y/s320/P7110009.JPG" width="320" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Tween 20-10 mL</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiQNQoKR2Rak3Mf30Y15x0s7949fCSGpt4w6UKTgYA3-XP4XRqd87HZFvS4zrwsK-xIsWdDtteHmrfcTLr2C7v5_yZvF68wJEBKDmGM0TN69Ie61eYQAdR2l4l-Zjtuw_A84XccDwVAhQc/s1600/P7110010.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="240" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiQNQoKR2Rak3Mf30Y15x0s7949fCSGpt4w6UKTgYA3-XP4XRqd87HZFvS4zrwsK-xIsWdDtteHmrfcTLr2C7v5_yZvF68wJEBKDmGM0TN69Ie61eYQAdR2l4l-Zjtuw_A84XccDwVAhQc/s320/P7110010.JPG" width="320" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Adjusting the pH to an Acidic Level</td></tr>
</tbody></table>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEi9AkLaj0mWsULrcKBO9jt0_-S4DctTrBxS8evaY-VcLYUnBgKCa23Y1R_24UeyRRTUBatub-2MXf0FtQ8nhZR_brqyZXJ6Sz3SYewX9XWfIPZTIiT-GKYuiW0aOVlGdNBhXUiUlPHnDWU/s1600/P7110011.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="240" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEi9AkLaj0mWsULrcKBO9jt0_-S4DctTrBxS8evaY-VcLYUnBgKCa23Y1R_24UeyRRTUBatub-2MXf0FtQ8nhZR_brqyZXJ6Sz3SYewX9XWfIPZTIiT-GKYuiW0aOVlGdNBhXUiUlPHnDWU/s320/P7110011.JPG" width="320" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Adjusting pH</td></tr>
</tbody></table>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEitI8w8-xPYRaG7hQdQmj8pVENHgLaCyjXDdywRIuJEr5yfrlYhzESDoqNuV5mm966WlJxTmHZKrHbRVytbFAwkaD_tTtA3fIvn6S51evDM2Q47RQJ268cTk_LYdYUGki2dBwOffoMyH-g/s1600/P7110012.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="320" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEitI8w8-xPYRaG7hQdQmj8pVENHgLaCyjXDdywRIuJEr5yfrlYhzESDoqNuV5mm966WlJxTmHZKrHbRVytbFAwkaD_tTtA3fIvn6S51evDM2Q47RQJ268cTk_LYdYUGki2dBwOffoMyH-g/s320/P7110012.JPG" width="240" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">The Finished Product</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEht-2YJvBU5TKRbjGTnG2c1He0T6rZkYBmlgh9P8n40zLRjSIQPl9uPowrZ65KESzz4B1m43wS9IUnfSqBE-CbIUIEo8X186joPyIUAQ61hT_XxRILcXdeHoN5B7SbPP3CSQ59GdFL8BUo/s1600/P7110013.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="320" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEht-2YJvBU5TKRbjGTnG2c1He0T6rZkYBmlgh9P8n40zLRjSIQPl9uPowrZ65KESzz4B1m43wS9IUnfSqBE-CbIUIEo8X186joPyIUAQ61hT_XxRILcXdeHoN5B7SbPP3CSQ59GdFL8BUo/s320/P7110013.JPG" width="240" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Aluminum Foil used to Protect Against Light Sensitivity </td></tr>
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<tr><td style="text-align: center;"><a href="https://static.thermoscientific.com/images/F57429~wl.jpg" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" src="https://static.thermoscientific.com/images/F57429~wl.jpg" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Commercial Stripping Buffer</td></tr>
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<span style="background-color: white;">So, instead of spending $134 to buy a commercial buffer, why not recreate it on your own? :)</span></div>
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<span style="background-color: white;">Other activities this week included preparing transfer buffer, running buffer, developing Western blots, transferring tissue to histology cassettes, and autoclaving. </span></div>Lauren Tolathttp://www.blogger.com/profile/05812297914953765766noreply@blogger.com0tag:blogger.com,1999:blog-8500808137876637614.post-82786759772432849712012-07-06T12:39:00.001-07:002012-07-06T13:29:29.158-07:00Week 7-The Research Proposal<div>
I hope everyone had a wonderful Independence Day! </div>
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This week has been action-packed, tension-filled, and stressful! My graduate student mentor, Qiying Fan, is ambitiously preparing for her research proposal and defense that she will soon present to a committee. This is extremely important because it partially defines her Ph.D. candidacy and she must prepare for a variety of questions to be thrown at her--including those from specialized experts hailing from Houston's medical district. Everyone has been trying to help out with her experiments as much as possible in order to allow her adequate preparation time. For practice, she will be presenting her proposal and defense to me. Her presentation includes a lot of information about the initiation and procession of atherosclerosis and the necessity to continue developing the relationship between a particular protein and the onset of atherosclerosis. It also includes a lot of information about the angiotensin II signaling pathway and A Kinase Anchoring proteins. Dr. McConnell told me to critique her analysis and her explanations about her presentation so that she will be better prepared when the real time comes!<br />
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One thing that Fan's research presentation showed me was the importance of having a strong connection between the background and the preliminary data. The background indicates the significance of the problem to be examined in the research project while the preliminary data targets a specific aspect of the problem. In this specific situation, Fan needed to make a stronger connection between her notes about atherosclerosis and the protein that she is investigating in the signal transduction pathway.<br />
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Additionally, two graduate students from TSU have been touring the facility lately. Dr. McConnell said that they are prospective lab members and are in the process of learning specific lab protocols/techniques. I was asked to introduce these students to what I've learned over the course of my stay here in the lab, including: autoclaving, filtration, and Western blots. How exciting is that? Just a few weeks ago I was in the process of being trained, and now, I get the opportunity to instruct. It's such a rewarding feeling.</div>
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This week I participated in developing Western blots, transferring heart tissue into histology cassettes, and preparing buffers for cardiomyocyte isolation. One buffer I had a fun time making was the Fura 2-AM loading buffer (cardiomyocyte perfusion buffer). It was difficult making this buffer because the substrates do not fully dissolve in the solution unless the pH is adjusted. The contents make the solution basic so hydrochloric acid needs to be added in order to reach proper homogeneity. Fura 2-AM is an intracellular calcium indicator that is ratiometric and UV light excitable. At low concentrations of the indicator, accurate measurements of the intracellular <span style="background-color: white;">Ca</span><sup style="background-color: white;">2+ </sup><span style="background-color: white;">concentration are able to be measured. </span><span style="background-color: white; vertical-align: super;"> </span></div>
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<tr><td class="tr-caption" style="text-align: center;">Cardiomyocyte Buffer-Preparation Sheet</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgd0zc4GGOrAwCKHVWkecBjE0HQutFPEGlFqUqbFfVtTpH-ZBRwySzP3yF9wQTQYFovVa8fSLTQXI1YcPXQho_8H0SN-JXWmTmToLSCL8ThyhEtvKDHYmjj8MQ5FV0tn2s6FARfou2mEUI/s1600/P7040022.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="200" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgd0zc4GGOrAwCKHVWkecBjE0HQutFPEGlFqUqbFfVtTpH-ZBRwySzP3yF9wQTQYFovVa8fSLTQXI1YcPXQho_8H0SN-JXWmTmToLSCL8ThyhEtvKDHYmjj8MQ5FV0tn2s6FARfou2mEUI/s200/P7040022.JPG" width="150" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Cardiomyocyte Buffer-Preparation</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiUJqqBgGzvk-PIHCx87kcTZs6BmGbYpzJyGAFzrz-c1MYRAsHYNA9RlDtFgRjbnVN3LqXnDCHm2Vqi2SzG7x0YBfQ_2fFh9SfDIzxR1288u4-LJuRhDtFm5FS2X5Fd-JJs36uha5Ng9jw/s1600/P7040025.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiUJqqBgGzvk-PIHCx87kcTZs6BmGbYpzJyGAFzrz-c1MYRAsHYNA9RlDtFgRjbnVN3LqXnDCHm2Vqi2SzG7x0YBfQ_2fFh9SfDIzxR1288u4-LJuRhDtFm5FS2X5Fd-JJs36uha5Ng9jw/s200/P7040025.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Sonal Performing Cardiomyocyte Isolation</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEg0O7JpH8Xq9_grHFlPFrjUcyGL9LclM1etn2PgyrV9LhEXg_lHMjaGsBVBWOZJk3NySYE3vnHIWfxQ6LT_tCYv32uW6D9EY6OBnzZIg4nHzZg20WIq2eE643BD97fosCR-BZJ62qvxqT8/s1600/P7050026.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="200" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEg0O7JpH8Xq9_grHFlPFrjUcyGL9LclM1etn2PgyrV9LhEXg_lHMjaGsBVBWOZJk3NySYE3vnHIWfxQ6LT_tCYv32uW6D9EY6OBnzZIg4nHzZg20WIq2eE643BD97fosCR-BZJ62qvxqT8/s200/P7050026.JPG" width="150" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Solution-Making</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEg_hxNbgFFidNbH8KrGfj7hnFSxslbOJF3bk38jlg9Q3V1amkPSNZ2PJWesffM5AURqQEgBsl47Bqiqc48zJms7r1EOqX6uxwqoreOwk8itot8fanLwJZ4ORUxifmiPRDm5GWpCtoR9W3A/s1600/P7050028.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEg_hxNbgFFidNbH8KrGfj7hnFSxslbOJF3bk38jlg9Q3V1amkPSNZ2PJWesffM5AURqQEgBsl47Bqiqc48zJms7r1EOqX6uxwqoreOwk8itot8fanLwJZ4ORUxifmiPRDm5GWpCtoR9W3A/s200/P7050028.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Solution-Making</td></tr>
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<table align="center" cellpadding="0" cellspacing="0" class="tr-caption-container" style="margin-left: auto; margin-right: auto; text-align: center;"><tbody>
<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhgPTuEn79aHeVCIZTFgRU5N_kIylyGiPIcxh3HqdHqnBd-peswyS_klSECLU30SYgqDeoZtGB5_c7ojJfVQI-eVT0eiFZFx1zo5HRG_nZudQvfqR6l_vAYDeszYPF3sMRQOaz_xHypogA/s1600/P7050033.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="200" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhgPTuEn79aHeVCIZTFgRU5N_kIylyGiPIcxh3HqdHqnBd-peswyS_klSECLU30SYgqDeoZtGB5_c7ojJfVQI-eVT0eiFZFx1zo5HRG_nZudQvfqR6l_vAYDeszYPF3sMRQOaz_xHypogA/s200/P7050033.JPG" width="150" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Cori and I</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEil5Rfot5KCoOMNMhQpuLumVjEKzE1MCo-4E8YVLQQ8ykJWhb2ps-i_4plXbY1aYwrUicB97unlsayEyprJs0r7dE0eHCE_kqt4yiMOl2BbJKFaDDgxxJ3abr6TCPPKB-poH11y4ie0gOY/s1600/P7050031.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="200" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEil5Rfot5KCoOMNMhQpuLumVjEKzE1MCo-4E8YVLQQ8ykJWhb2ps-i_4plXbY1aYwrUicB97unlsayEyprJs0r7dE0eHCE_kqt4yiMOl2BbJKFaDDgxxJ3abr6TCPPKB-poH11y4ie0gOY/s200/P7050031.JPG" width="150" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Solution Making</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhtm7MhggJ6lbZ5eF72eZHRtinUBc2aIziHljw9ToiCnXwUyawNlXKyMQN6nzbzn6AM9zLn2hkpyK1gTzeVL6RT73JIP2aS5fOOFyjYC6gkFIIV0BzT-47wb7Zw7xKfdkfnQrmOJN09sU0/s1600/P7050035.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhtm7MhggJ6lbZ5eF72eZHRtinUBc2aIziHljw9ToiCnXwUyawNlXKyMQN6nzbzn6AM9zLn2hkpyK1gTzeVL6RT73JIP2aS5fOOFyjYC6gkFIIV0BzT-47wb7Zw7xKfdkfnQrmOJN09sU0/s200/P7050035.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Adjusting the pH</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhhkqhlTyVS2JoCWALf38KBS2jLERt4xHBy6NFnckfQPS0_9Jkf7CFJL9BzP7cB8en4kukQZZXCP18Oe5xfKFDbhtTbMlHjYCai7ZAhEksUc1slakPK7t7PohFzJWVvBzPvIxIsPHXhY9w/s1600/P7050036.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhhkqhlTyVS2JoCWALf38KBS2jLERt4xHBy6NFnckfQPS0_9Jkf7CFJL9BzP7cB8en4kukQZZXCP18Oe5xfKFDbhtTbMlHjYCai7ZAhEksUc1slakPK7t7PohFzJWVvBzPvIxIsPHXhY9w/s200/P7050036.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Cori :)</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjKv6Fiz4KuIVUSSff0SEDuait6lYwaqR3eRVxf6YYxvEF7PYkWbnka1bL06LkBXlDmDJhRGH0uPtCQwaqjBkI8pbusd6s9WE2sZalpWLY_-Bfn7-nKMqQe3AaCpLW8bUkVEAxVOkJaaeo/s1600/P7050039.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="200" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjKv6Fiz4KuIVUSSff0SEDuait6lYwaqR3eRVxf6YYxvEF7PYkWbnka1bL06LkBXlDmDJhRGH0uPtCQwaqjBkI8pbusd6s9WE2sZalpWLY_-Bfn7-nKMqQe3AaCpLW8bUkVEAxVOkJaaeo/s200/P7050039.JPG" width="150" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">The Finished and Filtered Buffer!</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgrW-xgd4XRUGTh9s9LP3mHGIExRhi05utLTR9uc840t_y7fqWEoWH4N1XaABFaZpIMpzRl_6-c-QEdlsQYzQwiV4DULSKET_FMydcav4nEHFV6Ye8KKzAly6FhCjm2ywhcRpy1azBKa40/s1600/P7050040.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgrW-xgd4XRUGTh9s9LP3mHGIExRhi05utLTR9uc840t_y7fqWEoWH4N1XaABFaZpIMpzRl_6-c-QEdlsQYzQwiV4DULSKET_FMydcav4nEHFV6Ye8KKzAly6FhCjm2ywhcRpy1azBKa40/s200/P7050040.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Fan Preparing Her Research Proposal</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhfoq0WFNikkxxQONRgfZajnZZ_ynmjg5I9n4NiQrMgf4aHX_05PwmgOQGmitfuLwk0QUEmza8b1mXVXvCWgSLcTBYQxzUUurgdtaJF7kX-vEckllbhL8MrBz5Io2SY1nH6aRgeJaNZBKo/s1600/P7060042.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhfoq0WFNikkxxQONRgfZajnZZ_ynmjg5I9n4NiQrMgf4aHX_05PwmgOQGmitfuLwk0QUEmza8b1mXVXvCWgSLcTBYQxzUUurgdtaJF7kX-vEckllbhL8MrBz5Io2SY1nH6aRgeJaNZBKo/s200/P7060042.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Fan-Practicing Her Presentation</td></tr>
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</div>Lauren Tolathttp://www.blogger.com/profile/05812297914953765766noreply@blogger.com0tag:blogger.com,1999:blog-8500808137876637614.post-9885685677867008192012-06-29T12:42:00.002-07:002012-06-29T13:02:48.688-07:00Week 6-Seeking the Bigger Picture & Meeting the Team<div class="separator" style="clear: both; text-align: justify;">
<span style="background-color: white; text-align: left;">After hearing about a variety of research projects that are underway and have been completed on this campus, I am now even more excited about continuing my ongoing project on heart disease. With the legitimate concerns about pandemics of AIDS and behavioral disorders, it's easy to forget that cardiovascular disease kills more people each year worldwide than any other disease. We've grown so accustomed to hearing about heart disease that we even consider it a natural cause of death and tend to disregard a comprehensive solution to an obvious growing issue. I can even attest to the effects of heart disease, as it has personally plagued loved ones and family friends. Some of the most recent well-publicized deaths are even linked to such: Whitney Houston suffered from atherosclerosis, a disease of the arteries as a result of fatty materials such as cholesterol. The best part about joining this lab and its ongoing project is the fact that I feel like I am tackling two developments at the same time--discovering the role of a particular protein in heart disease and finding a solution to modulate this protein and, perhaps, a solution to heart disease in general through pharmacodynamics. I am ecstatic that this isn't just a project that stops at the discovery of a particular thing--it seeks to improve the human condition through application.</span></div>
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Even more exciting is the fact that one of my research team members, Ashley, is on the brink of submitting a huge paper that includes data/data analysis dating back (whew..a mouthful) to 2010! A lot of what she is including in her paper predates the contributions I have made since joining the lab; however, the implications of a successful publication are enormous for the rest of the lab work to follow.<br />
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Speaking of my research team, allow me to introduce you to some of them! They have been a part of Dr. McConnell's lab for as long as he has been a PI, and I can assure you they are a TREMENDOUS help (and motivation) to my research experience:<br />
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<a href="https://sphotos.xx.fbcdn.net/hphotos-snc6/216802_171046782966679_6220678_n.jpg" imageanchor="1" style="clear: left; float: left; margin-bottom: 1em; margin-right: 1em;"><img border="0" height="150" src="https://sphotos.xx.fbcdn.net/hphotos-snc6/216802_171046782966679_6220678_n.jpg" width="200" /></a><b><u>Name</u></b>: Cori Wijaya<br />
<span style="font-family: 'times new roman', 'new york', times, serif;"><b><u>Education</u></b>: B.S. in Biochemistry-University of Houston</span><br />
<span style="font-family: 'times new roman', 'new york', times, serif;"><b><u>What sparked her interest in research</u></b>: </span><span style="background-color: white;"><span style="font-family: 'times new roman', 'new york', times, serif;">She started doing research work during one summer in her undergraduate senior year. She found that doing hands-on work in the lab helped her with her studying: what she read in her textbooks actually started to make sense! As she graduated and started working in the lab, she found it exciting to do experiments that helped to elucidate the questions and hypothesis of the research work.</span></span><br />
<span style="background-color: white;"><span style="font-family: 'times new roman', 'new york', times, serif;"><b><u>The best part about her experience in Dr. McConnell's lab</u></b>: Getting to know people who share her knowledge and skills</span></span><br />
<span style="font-family: 'times new roman', 'new york', times, serif;"><b><u>Advice she gave me about continuing to pursue research:</u></b> </span><span style="background-color: white;"><span style="font-family: 'times new roman', 'new york', times, serif;">Keep a curious and open mind. Research is a long process with so many little things to take care of-but be glad that you are a part of it. Spend a long enough time in one lab to learn something, but also give yourself a chance to try a different lab and see how other labs work differently.</span></span><br />
<span style="font-family: 'times new roman', 'new york', times, serif; font-size: x-small;"><br /></span><br />
<a href="https://sphotos.xx.fbcdn.net/hphotos-snc7/304188_10150372033471568_110912392_n.jpg" imageanchor="1" style="clear: left; float: left; margin-bottom: 1em; margin-right: 1em;"><img border="0" height="200" src="https://sphotos.xx.fbcdn.net/hphotos-snc7/304188_10150372033471568_110912392_n.jpg" width="120" /></a><b><u>Name</u></b>: Abeer Rababa'h<br />
<span style="font-family: 'times new roman', 'new york', times, serif;"><b><u>Education</u></b>: </span><span style="background-color: white;"><span style="font-family: 'times new roman', 'new york', times, serif;">Master degree in Clinical Pharmacology/ Jordan University of Science and Technology</span></span><br />
<span style="font-family: 'times new roman', 'new york', times, serif;"><b><u>What sparked her interest in research</u></b>: </span><span style="background-color: white;"><span style="font-family: 'times new roman', 'new york', times, serif;">She has an interest in correlating and interpreting data into biological facts that help downstream in achieving new clinical aspects.</span></span><br />
<span style="background-color: white;"><span style="font-family: 'times new roman', 'new york', times, serif;"><b><u>The best part about her experience in Dr. McConnell's lab</u></b>: </span></span><span style="background-color: white;"><span style="font-family: 'times new roman', 'new york', times, serif;">Research with Dr. McConnell gives her the opportunity to interpret many findings in a logical way, find explanations for every single piece of data, convert bored data into exciting findings, look for something from a different point of view in a comfortable friendly research environment.</span></span><br />
<span style="font-family: 'times new roman', 'new york', times, serif;"><b><u>Advice she gave me about continuing to pursue research:</u></b> </span><span style="background-color: white;"><span style="font-family: 'times new roman', 'new york', times, serif;">My advice is to get as much experience as you can from the lab. Every small contribution may participate in a big achievement in your career goal. Challenge yourself and be patient. Being hard working is a requirement for the research road.</span></span><br />
<span style="font-family: 'times new roman', 'new york', times, serif; font-size: x-small;"><br /></span><br />
<a href="https://sphotos.xx.fbcdn.net/hphotos-ash3/18668_240055098287_924804_n.jpg" imageanchor="1" style="clear: left; float: left; margin-bottom: 1em; margin-right: 1em;"><img border="0" height="200" src="https://sphotos.xx.fbcdn.net/hphotos-ash3/18668_240055098287_924804_n.jpg" width="153" /></a><b><u>Name</u></b>: Andrea Diaz Diaz<br />
<span style="font-family: 'times new roman', 'new york', times, serif;"><b><u>Education</u></b>: </span><span style="background-color: white;"><span style="font-family: 'times new roman', 'new york', times, serif;">B.S. in Biology and Biochemistry-University of Houston</span></span><br />
<span style="font-family: 'times new roman', 'new york', times, serif;"><b><u>What sparked her interest in research</u></b>: She</span><span style="background-color: white;"><span style="font-family: 'times new roman', 'new york', times, serif;"> took some classes in college with a wonderful teacher that motivated her to do research.</span></span><br />
<span style="background-color: white;"><span style="font-family: 'times new roman', 'new york', times, serif;"><b><u>The best part about her experience in Dr. McConnell's lab</u></b>: She</span></span><span style="background-color: white;"><span style="font-family: 'times new roman', 'new york', times, serif;"> gets to do surgeries (on mice, that is!)</span></span><br />
<span style="font-family: 'times new roman', 'new york', times, serif;"><b><u>Advice she gave me about continuing to pursue research:</u></b> </span><span style="background-color: white;"><span style="font-family: 'times new roman', 'new york', times, serif;">It is a very interesting career where you can learn many things about any topic you are interested in!</span></span><br />
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEib9z9Q4N-wQmPxMtkHJF7hVSKTe7vzjFmkZ9KI8_zIPOq-fAC7gt0A4-h93EDeoESTE2ngobWKKjd9Auj0T63Mlrxk-D5-Wp6yKPV81oil2s743sVUl6peF0bTc46VtU5z7D30iRautbE/s1600/P6250001.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="200" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEib9z9Q4N-wQmPxMtkHJF7hVSKTe7vzjFmkZ9KI8_zIPOq-fAC7gt0A4-h93EDeoESTE2ngobWKKjd9Auj0T63Mlrxk-D5-Wp6yKPV81oil2s743sVUl6peF0bTc46VtU5z7D30iRautbE/s200/P6250001.JPG" width="150" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">X-Ray Film Processor</td></tr>
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<table align="center" cellpadding="0" cellspacing="0" class="tr-caption-container" style="margin-left: auto; margin-right: auto; text-align: center;"><tbody>
<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjHjpSfC6WUNt6qpfexFw2T815rHPlsIT_5fQz5lcneA1kuU8Y1XKCCiKMpw6xEILl6GIQ2kc38wXA9bHB5yOW6-jTelKD8aB6G4ZsLHckvSwfsb1trtDnXbbuBAUTHxLCbF_HsAHLHJG4/s1600/P6250002.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjHjpSfC6WUNt6qpfexFw2T815rHPlsIT_5fQz5lcneA1kuU8Y1XKCCiKMpw6xEILl6GIQ2kc38wXA9bHB5yOW6-jTelKD8aB6G4ZsLHckvSwfsb1trtDnXbbuBAUTHxLCbF_HsAHLHJG4/s200/P6250002.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Developing the Film-Western Blot</td></tr>
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<table align="center" cellpadding="0" cellspacing="0" class="tr-caption-container" style="margin-left: auto; margin-right: auto; text-align: center;"><tbody>
<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgzOIGl10zZ7qvSJqOPaY3JZTAJQ4YsT5Z4TNyonvqAHsrxMnt44XJcah5WQUlKd1qUM0fc5-TAqwhvtVR1qNvotBWDEratrYBEiZ3QCOPg4D8YrUyxVeC2A7l3OBeqU88Ek6ogigLPcG8/s1600/P6250003.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgzOIGl10zZ7qvSJqOPaY3JZTAJQ4YsT5Z4TNyonvqAHsrxMnt44XJcah5WQUlKd1qUM0fc5-TAqwhvtVR1qNvotBWDEratrYBEiZ3QCOPg4D8YrUyxVeC2A7l3OBeqU88Ek6ogigLPcG8/s200/P6250003.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Labeling the Film</td></tr>
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<table align="center" cellpadding="0" cellspacing="0" class="tr-caption-container" style="margin-left: auto; margin-right: auto; text-align: center;"><tbody>
<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEizhfPTz9VGA_BlrXndqPneLeNInk1YAVN893EeEgPbLbII4d7AVnbOfKAroLOCVzXBnpAuCVA8jqCRpEqa66W0Jq1DFLhBNsZj_4X4HhUyAS6-tIMXbJ23jl8MMFI5ELFh2MxTw5N6QVM/s1600/P6260004.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEizhfPTz9VGA_BlrXndqPneLeNInk1YAVN893EeEgPbLbII4d7AVnbOfKAroLOCVzXBnpAuCVA8jqCRpEqa66W0Jq1DFLhBNsZj_4X4HhUyAS6-tIMXbJ23jl8MMFI5ELFh2MxTw5N6QVM/s200/P6260004.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Discovery Workshop-Using EndNote for your References</td></tr>
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<table align="center" cellpadding="0" cellspacing="0" class="tr-caption-container" style="margin-left: auto; margin-right: auto; text-align: center;"><tbody>
<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEivpnBv0tHxTqkj7A9cvSt3pduhB4YrDU7pB-o-uswbeoh9QVCqx7zC34Srxc417glOJKnyPAJfIXtLv9oKXai_5pyquKVV2wbN_YLahAq3UvUG9Ey7SLpY6A7gpjVeZMDcSQslME8PDZw/s1600/P6260005.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEivpnBv0tHxTqkj7A9cvSt3pduhB4YrDU7pB-o-uswbeoh9QVCqx7zC34Srxc417glOJKnyPAJfIXtLv9oKXai_5pyquKVV2wbN_YLahAq3UvUG9Ey7SLpY6A7gpjVeZMDcSQslME8PDZw/s200/P6260005.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Program: EndNote</td></tr>
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<table align="center" cellpadding="0" cellspacing="0" class="tr-caption-container" style="margin-left: auto; margin-right: auto; text-align: center;"><tbody>
<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiXBngfjN9h4GigagYOJkNwjJm4BY2cBQXxSvu1tPw-34KhpE214xcWnIb4ozXfUkhkU5vCiy3LGd5eVFm84wRMlyIyyzV543tBhDgGOc5DtdMOgJ4kn93o3v1VACeTK63YAQTg6rglJc4/s1600/P6290006.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiXBngfjN9h4GigagYOJkNwjJm4BY2cBQXxSvu1tPw-34KhpE214xcWnIb4ozXfUkhkU5vCiy3LGd5eVFm84wRMlyIyyzV543tBhDgGOc5DtdMOgJ4kn93o3v1VACeTK63YAQTg6rglJc4/s200/P6290006.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Heart Tissue Stored in Ethanol</td></tr>
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<table align="center" cellpadding="0" cellspacing="0" class="tr-caption-container" style="margin-left: auto; margin-right: auto; text-align: center;"><tbody>
<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgFewbsNegBKRWhdPTaNHCTqY80wUtLbXE8OBphogkwtmD5Y2Z-Bcv8yzCOVletcBlgOXSEJbOEtaBxHdxRRHK_xIipIN63p__E2mncRtgFUpqpLyDvkQFnFlhg4DEO5NasOZbNZqwnr8Q/s1600/P6290007.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgFewbsNegBKRWhdPTaNHCTqY80wUtLbXE8OBphogkwtmD5Y2Z-Bcv8yzCOVletcBlgOXSEJbOEtaBxHdxRRHK_xIipIN63p__E2mncRtgFUpqpLyDvkQFnFlhg4DEO5NasOZbNZqwnr8Q/s200/P6290007.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Collected Heart Tissue</td></tr>
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<table align="center" cellpadding="0" cellspacing="0" class="tr-caption-container" style="margin-left: auto; margin-right: auto; text-align: center;"><tbody>
<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhbxN8hyphenhyphenqKcWXUTKilUC1im6fnPz2p6-l-N4S-JroHJBBcCOc3-t7S4M97998o3VVXcAyUNQjAXVTchsGKf6m9zwX9ZVwKw8mC4S_M9mKQf5t_XSmZVvUQTbSyMQAAcFqW9wVHeIg8sQ-Q/s1600/P6290011.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="200" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhbxN8hyphenhyphenqKcWXUTKilUC1im6fnPz2p6-l-N4S-JroHJBBcCOc3-t7S4M97998o3VVXcAyUNQjAXVTchsGKf6m9zwX9ZVwKw8mC4S_M9mKQf5t_XSmZVvUQTbSyMQAAcFqW9wVHeIg8sQ-Q/s200/P6290011.JPG" width="150" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Moving Tissue to Histology Cassettes</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgoPxTq-2RJOPXL4vcEGEvAU9d_e5AGOx_hR2ADQHTjsQwAe5Soh4HwRdz2LiEALm1WIfqOqfMzwFUX_CpeiDmuyM4lZxn24KqvP-5UCNhAvFD7gP0L5s4kkUroAzEqTe3zF-iTVlDTeK8/s1600/P6290012.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="200" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgoPxTq-2RJOPXL4vcEGEvAU9d_e5AGOx_hR2ADQHTjsQwAe5Soh4HwRdz2LiEALm1WIfqOqfMzwFUX_CpeiDmuyM4lZxn24KqvP-5UCNhAvFD7gP0L5s4kkUroAzEqTe3zF-iTVlDTeK8/s200/P6290012.JPG" width="150" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Moving Tissue to Histology Cassettes</td></tr>
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<br />Lauren Tolathttp://www.blogger.com/profile/05812297914953765766noreply@blogger.com0tag:blogger.com,1999:blog-8500808137876637614.post-15351528465322622312012-06-22T14:23:00.000-07:002012-06-25T20:26:48.779-07:00Week 5-Western Blots Galore!My primary focus this week was continuing the development of various Western Blots. One of the members of our research team, Abeer, came back this week from maternity leave to continue her project. The lab was especially busy this week because a lot of buffer solutions had to be prepared in order to practice the cardiomyocyte isolations that will begin next week.<br />
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One thing I really enjoy about this research experience is learning (and practicing) various biological lab techniques that was only briefly mentioned in my biology class. Before entering the lab, I had only heard of a "Southern blot." This technique is used to detect a specific DNA sequence in DNA samples and uses probe hybridization. Even though the names are similar, a Western blot is completely different. It targets the identification of specific proteins in a given sample through gel electrophoresis and antibody hybridization.<br />
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The first step in a Western blot is to prepare the tissue using a sonicator (there are various techniques in preparing the tissue, but the primary one used in the lab is with a sonicator). The sonicator breaks down the solid cell wall of the cells in the particular sample and enables the protein found within the cell to be freed.<br />
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The next step is to perform gel electrophoresis. This separates the proteins of the sample by electric charge and molecular weight. Smaller proteins move faster through the gel and the proteins are separated by size. The proteins move through a specific agar gel; the more concentrated the agar gel is, the better the resolution of the lower molecular weight proteins. (agar gel is created using a powder, diluent, and heat - similar to jello!) Conversely, the lower the concentration, the better the resolution of higher molecular weight proteins. The samples are loaded into walls in the gel, with one lane reserved for a marker. This marker is a commercially made mixture of proteins having pre-determined molecular weights. They are also stained to produce highly visible and colored bands.<br />
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<table align="center" cellpadding="0" cellspacing="0" class="tr-caption-container" style="margin-left: auto; margin-right: auto; text-align: center;"><tbody>
<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhSCqOgZw2q2NgS-0KI5GI0kZMGRBsHCX258GTX6OGuL4Sd4Qrz49D_ERc4S2UoVZ4v47Wy67YnKHSUfQ_rclPnUUKnbYAOjGIqHdCKFVFTbc0VMwot0UiYrCOCZaPV2SjV7ssYC1XUAfM/s1600/P6220029.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="200" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhSCqOgZw2q2NgS-0KI5GI0kZMGRBsHCX258GTX6OGuL4Sd4Qrz49D_ERc4S2UoVZ4v47Wy67YnKHSUfQ_rclPnUUKnbYAOjGIqHdCKFVFTbc0VMwot0UiYrCOCZaPV2SjV7ssYC1XUAfM/s200/P6220029.JPG" width="150" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Running Buffer</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEia82gxSBI1PU6ppYDsa6nNDpSm_7jKhWHWzZ_xjSQv604INC8SLof3JYmlQcUwOi-JVB_vWlyjhrnV3kvP61zYztzPb-MhriY0cIaPwcCqYEOIPF3pOh1Gj9j3CXYVA5a8dNgbB69kGHQ/s1600/P6220030.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEia82gxSBI1PU6ppYDsa6nNDpSm_7jKhWHWzZ_xjSQv604INC8SLof3JYmlQcUwOi-JVB_vWlyjhrnV3kvP61zYztzPb-MhriY0cIaPwcCqYEOIPF3pOh1Gj9j3CXYVA5a8dNgbB69kGHQ/s200/P6220030.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Samples to be Loaded</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEi0FjzBK-q-fl6BFemU1-Ndli8h9FLqJFM2eLxtTIO7jMZap2H1v_9mTFnTHZTSxSpbajEhdaoP7Kj3uSapXOxF_Wwso-4hyphenhyphenRadKdMkwrx6pXfQsNZT15WgXb1my7cBSBTRG1JMGFgj5dw/s1600/P6220033.JPG" imageanchor="1" style="clear: right; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEi0FjzBK-q-fl6BFemU1-Ndli8h9FLqJFM2eLxtTIO7jMZap2H1v_9mTFnTHZTSxSpbajEhdaoP7Kj3uSapXOxF_Wwso-4hyphenhyphenRadKdMkwrx6pXfQsNZT15WgXb1my7cBSBTRG1JMGFgj5dw/s200/P6220033.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Loading Samples</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhsKXSwgujClNQxOwPxtOO9olN4JAICP376T2qHfO5OGX3_vWrwIjVH1JpF9_MWHUTN42aalSFnYPj5OzlEqENgp7k4hUU4Z1JlLdaogUO5pRFDhCPJaZXpiySFSjMSzxG3VH2PCKiWqlM/s1600/P6220031.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhsKXSwgujClNQxOwPxtOO9olN4JAICP376T2qHfO5OGX3_vWrwIjVH1JpF9_MWHUTN42aalSFnYPj5OzlEqENgp7k4hUU4Z1JlLdaogUO5pRFDhCPJaZXpiySFSjMSzxG3VH2PCKiWqlM/s200/P6220031.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Loading Samples</td></tr>
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<tr><td class="tr-caption" style="text-align: center;">Electrophoresis with an Electric Current</td></tr>
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After running the gel, the proteins are transferred to what is called a membrane (PVDF paper). The primary method for transferring the proteins to the membrane is a process known as electroblotting. This uses an electric current to to pull proteins from the gel into the PVDF membrane. What I find really unique about this process is that the electric current pulls the proteins from the gel onto the membrane in the exact same order and size as on the gel! As a result of this process, the proteins are exposed on a surface and this is used for detection.<br />
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Next, steps must be taken to prevent the antibody from binding to a non-target protein. This technique is called blocking and it involves using TBST (Tris-Buffered Saline with Tween 20). The protein in the dilute solution attaches to the membrane in all places where the target proteins have not attached. When the antibody is added, there is no room for it to attach other than on the binding sites of the specific target protein.<br />
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Detection may be perhaps the most important step in the entire process and is when antibody hybridization occurs. In my lab, we practice this in two steps: a primary and secondary antibody. The PVDF membrane is probed with a particular antibody (mouse antibody, rabbit antibody, donkey antibody, etc.). When a match occurs, a different color on the membrane is produced.<br />
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Last, the membrane is incubated with ECL (a solution used in chemiluminescence) which allows for chemiluminescent detection. The light is detected by a photographic film and results are printed.<br />
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Other activities this week included autoclaving glass bottles to fill them with buffers and practicing cardiomyocyte isolations in order to adequately perform next week's activities.<br />
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjpWbgTHtuXwR7xWKad6tUw1WEtAlEcHPBVJ0qpzxE3pt3Nd6BpRw4UllOIxDG8LdMerbhHaNoIsxR9b_iixGyQxKcqxo3_CVe-8UlamBIX6Z3lYkRmH6uPIHjyqXabXBd1-OF3fr_4J84/s1600/P6180018.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="200" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjpWbgTHtuXwR7xWKad6tUw1WEtAlEcHPBVJ0qpzxE3pt3Nd6BpRw4UllOIxDG8LdMerbhHaNoIsxR9b_iixGyQxKcqxo3_CVe-8UlamBIX6Z3lYkRmH6uPIHjyqXabXBd1-OF3fr_4J84/s200/P6180018.JPG" width="150" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Autoclave Unit</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiuxaCEtQJreiu2ugO8FnHQBGQqIdhAKjcYxDsmlQ-Wph1RuGnsHrja-C2SZ6e6WthaiVS1cLI5NJZbwHXVBeVi_qzmCpJ5N2re8MSzoLwHXxSY6EhLrYso43DWYWKhd8uBjwmWTTybWwg/s1600/P6180019.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiuxaCEtQJreiu2ugO8FnHQBGQqIdhAKjcYxDsmlQ-Wph1RuGnsHrja-C2SZ6e6WthaiVS1cLI5NJZbwHXVBeVi_qzmCpJ5N2re8MSzoLwHXxSY6EhLrYso43DWYWKhd8uBjwmWTTybWwg/s200/P6180019.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Autoclaved Bottles </td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEizPCi5PUSW0lfvv0_ZfDjulASsUmmL3SHp-eh0MAp5V8y_YBZklDvz7B6FusR0DlhYL9HBw6_9lhyphenhyphenPJrzMfL6TbG0hHAaTH98tWtMpOjUvSAy-P2iOH6RkBCHVQYK9-MMGUzPAMLs9kzg/s1600/P6220028.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEizPCi5PUSW0lfvv0_ZfDjulASsUmmL3SHp-eh0MAp5V8y_YBZklDvz7B6FusR0DlhYL9HBw6_9lhyphenhyphenPJrzMfL6TbG0hHAaTH98tWtMpOjUvSAy-P2iOH6RkBCHVQYK9-MMGUzPAMLs9kzg/s200/P6220028.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Cardiomyocyte Isolation</td></tr>
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<br />Lauren Tolathttp://www.blogger.com/profile/05812297914953765766noreply@blogger.com0tag:blogger.com,1999:blog-8500808137876637614.post-57280500658661335782012-06-15T18:05:00.000-07:002012-07-06T13:06:51.788-07:00Week 4-Dealing with Collected TissueAfter tons of paper-writing, peer editing, and data-analyzing from the previous week, this week required much more activity. Some of the mice from our experimental groups finally reached 21 weeks of age and were ready to get their blood pressure & heart values confirmed by an invasive procedure dealing with the carotid artery. With this procedure completed, it was time to carefully stain the collected aortas in order to conduct lipid lesion formation analysis.<br />
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First, I drained the aortas with a 100% propylene glycol solution. This dehydrates the tissue and prepares them for staining. Next, I incubated the aortas with the stain for 4 hours at room temperature. After the 4 hours were completed, I washed the aortas with 85% propylene glycol solution. Last, I incubated the aortas overnight with a PBS (saline) solution in order to preserve the tissue and prevent it from drying out.<br />
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEghU696IzomDRqw5u3WZjwTIKeiDguj3D628ENy6BCFfpGFlNm4gtyyiUqgdDmg4V9b3kzmevZl1Z6yf0fQ_Qa2CSjHj3jTUUUKcVaNP35XpI9hJRovjmm-t2PrG26kbWbY4osR4M1g9fk/s1600/P6110004.JPG" imageanchor="1" style="clear: right; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEghU696IzomDRqw5u3WZjwTIKeiDguj3D628ENy6BCFfpGFlNm4gtyyiUqgdDmg4V9b3kzmevZl1Z6yf0fQ_Qa2CSjHj3jTUUUKcVaNP35XpI9hJRovjmm-t2PrG26kbWbY4osR4M1g9fk/s200/P6110004.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Collected Tissue </td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEghZ0oXtmWDkIOumVwQIh9qa-zKbqjQ6qSteQkBF3PMCJcLYwYkKtQE23DqKw2jZ6CWxlJYvpixJ5fsl2dFvs34UqygpNtmhRhGMLYj2BjKvhRoofN5jrlUDvOP7CEv88tDTj1_6iXYNv8/s1600/P6110009.JPG" imageanchor="1" style="clear: left; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEghZ0oXtmWDkIOumVwQIh9qa-zKbqjQ6qSteQkBF3PMCJcLYwYkKtQE23DqKw2jZ6CWxlJYvpixJ5fsl2dFvs34UqygpNtmhRhGMLYj2BjKvhRoofN5jrlUDvOP7CEv88tDTj1_6iXYNv8/s200/P6110009.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">The Staining Process </td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiFUxAx0VQ4M6RTaXqs_DnwA10ly_kKiPcAK8Clr5qkC79065COWsw4vrWz-QsyueF-hFhUf8a3NIN6sWpDRfsh4sfSGzBmYjVp2S6OdzYJ9Fl2XLHV60I2VBz7Sy3V3ORYlZ1prpy0SQM/s1600/P6110007.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="200" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiFUxAx0VQ4M6RTaXqs_DnwA10ly_kKiPcAK8Clr5qkC79065COWsw4vrWz-QsyueF-hFhUf8a3NIN6sWpDRfsh4sfSGzBmYjVp2S6OdzYJ9Fl2XLHV60I2VBz7Sy3V3ORYlZ1prpy0SQM/s200/P6110007.JPG" width="150" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">100% Propylene Glycol</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgPbSJBkLAe1vowPJbKJzaHM0gMICYENdRyQngCIHuD8C2S8iszkEI-7AwcVkYumTDa3gn0849gXenndjxvIX3Cc0kCS8A3IVgZygY3eNuiRSg1HsE5rwC0aCI2N3v8vv6nUs8d6o6qcro/s1600/P6110006.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgPbSJBkLAe1vowPJbKJzaHM0gMICYENdRyQngCIHuD8C2S8iszkEI-7AwcVkYumTDa3gn0849gXenndjxvIX3Cc0kCS8A3IVgZygY3eNuiRSg1HsE5rwC0aCI2N3v8vv6nUs8d6o6qcro/s200/P6110006.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Collected Tissue</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEj-ZZ12qyfOpT274npkXqI6Rj5kYTPa_lezAM7IRNUBAXqPNaC5KfEPW6sE_L7YbRmvzAfbOOiXp57HkQy7VhVpPRX7nkqViVbQ_SPQxCfrlpmjR0QWZWuPvg9u0PILT-mr4XbvnhIhJtE/s1600/P6110005.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEj-ZZ12qyfOpT274npkXqI6Rj5kYTPa_lezAM7IRNUBAXqPNaC5KfEPW6sE_L7YbRmvzAfbOOiXp57HkQy7VhVpPRX7nkqViVbQ_SPQxCfrlpmjR0QWZWuPvg9u0PILT-mr4XbvnhIhJtE/s200/P6110005.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">PBS Solution</td></tr>
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In addition to this process, I learned how to filter the substrates to make the stain solution! This required a biosafety cabinet and a fume hood to catch the vapors of this potentially hazardous substance. This process also requires the use of a vacuum to help filter the substrate from the liquid. With a special device that connects to the top of the bottle, the vacuum acts on the substrate found in the upper portion of the device and drains all of the liquid to the bottom jar where it is then collected and stored. Because the stain is light-sensitive, we made sure to cover the bottle in aluminum foil to prevent the light from changing the chemical properties of the stain.<br />
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEheAu5Q_3Ay8QCuOq6QrOeo-WdfV6cUrZR5DCbWkND6LkOtzAQgoZINX6N5JasDQUgQ07DTxKfqAE5KfV7Z9fRNhc0qLjuyON1xZbwOOoZjgKGsXXsZQOiv3Vau4pAM0lF7giyjnA1YGRE/s1600/P6110008.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEheAu5Q_3Ay8QCuOq6QrOeo-WdfV6cUrZR5DCbWkND6LkOtzAQgoZINX6N5JasDQUgQ07DTxKfqAE5KfV7Z9fRNhc0qLjuyON1xZbwOOoZjgKGsXXsZQOiv3Vau4pAM0lF7giyjnA1YGRE/s200/P6110008.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Filtering Oil Red O</td></tr>
</tbody></table>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhZZErtkOmUskG4hMPrPixNCUS4HAbLYOW_x1W2O4G-87Ibvy3fQwL_IfT6JbZ8sdsel_bfmRTHKyZF03WUnSJ-09vh17w6df6lysXHcR773BTrhk7xfMtgwB7U-Vv7HN9NSv55F3qqgIY/s1600/P6110011.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="200" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhZZErtkOmUskG4hMPrPixNCUS4HAbLYOW_x1W2O4G-87Ibvy3fQwL_IfT6JbZ8sdsel_bfmRTHKyZF03WUnSJ-09vh17w6df6lysXHcR773BTrhk7xfMtgwB7U-Vv7HN9NSv55F3qqgIY/s200/P6110011.JPG" width="150" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Oil Red O Solution-Final Product</td></tr>
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Other activities this week included washing the membranes in ongoing Western Blots, switching out the PBS solution that was used to store the collected aortas, prepration of TBST & milk, and storage of membranes from Western Blots into sealed packs.<br />
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjUvnjANBRdzyh6tsqOfSoDMMOSVx_XlJSDy2OoS-4AEfYXIM6N8rXyO2ehGOgO81MIGL7F_kSPQjgAFkgC4_YZg_PhpOJynNFwkMYVf_cxDYirO2pD1KxdCgBX52bCBNCaruc7suhnvCM/s1600/P6120017.JPG" imageanchor="1" style="clear: right; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjUvnjANBRdzyh6tsqOfSoDMMOSVx_XlJSDy2OoS-4AEfYXIM6N8rXyO2ehGOgO81MIGL7F_kSPQjgAFkgC4_YZg_PhpOJynNFwkMYVf_cxDYirO2pD1KxdCgBX52bCBNCaruc7suhnvCM/s200/P6120017.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Washing the Membranes </td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgx5_DlFVPrmabJElq8oiUvyveuCSIsI6-pr3XBLWBaMSgcO4_0Eb7GVOuCtm1els3m10qQpZzWNJYc4rxnVLLQCKuh_ets9Nbp5caSqAlidtd_k4yD9KSWnOU0qdZkDzB1jUOd4IYv5sI/s1600/P6110001.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgx5_DlFVPrmabJElq8oiUvyveuCSIsI6-pr3XBLWBaMSgcO4_0Eb7GVOuCtm1els3m10qQpZzWNJYc4rxnVLLQCKuh_ets9Nbp5caSqAlidtd_k4yD9KSWnOU0qdZkDzB1jUOd4IYv5sI/s200/P6110001.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Incubating Western Blot</td></tr>
</tbody></table>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEha4GcPW21HMYo4HD96-mxY0_bBTA1yncgDLi8zopcJxSPJ3mvYAh62kWNFNteeJy_RozopEkjdfgOmBuSVzjrt691Se_OUf9GbZ5gUsPejzForim8vyFdP8t2buqYLCZE555AUmwIqdwo/s1600/P6120016.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEha4GcPW21HMYo4HD96-mxY0_bBTA1yncgDLi8zopcJxSPJ3mvYAh62kWNFNteeJy_RozopEkjdfgOmBuSVzjrt691Se_OUf9GbZ5gUsPejzForim8vyFdP8t2buqYLCZE555AUmwIqdwo/s200/P6120016.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Milk+Antibodies </td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjgIhIGoHHO8oOjJFQCcVxEZP-sOfSPHlRf8mp7CulYkRZ4bd3Y0qrSQ5kfa-vT2sfGecrN_olgRlTNwaEsIgSuYrp5M1fp4thsUzdI7NkApxZr1ZpeEFVD1gobFGinRiBBoEszJRZjp8I/s1600/P6110003.JPG" imageanchor="1" style="clear: left; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjgIhIGoHHO8oOjJFQCcVxEZP-sOfSPHlRf8mp7CulYkRZ4bd3Y0qrSQ5kfa-vT2sfGecrN_olgRlTNwaEsIgSuYrp5M1fp4thsUzdI7NkApxZr1ZpeEFVD1gobFGinRiBBoEszJRZjp8I/s200/P6110003.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Membrane from Western Blot</td></tr>
</tbody></table>
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</div>Lauren Tolathttp://www.blogger.com/profile/05812297914953765766noreply@blogger.com0tag:blogger.com,1999:blog-8500808137876637614.post-76462632489994851682012-06-08T13:39:00.000-07:002012-06-25T20:28:31.300-07:00Week 3-A Very Happy Birthday and Some pH-un!<div class="separator" style="clear: both; text-align: center;">
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This week came to prove that a phenomenal research team and a great work environment really do make an opportunity like this worthwhile. Aside from observing and performing lab experiments, there's a lot of thinking and collaborating. Every person on the research team is examining something different, but at the end of the day, everybody's individual parts are brought together to answer a broader question. Additionally, while research may require a lot of patience and thinking, there is definitely a "fun" dimension involved.<br />
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In the process of blogging about my team's specific project details, it's necessary to be wary of what I post. This is because if too much information is publically posted about a particular experiment or our overall research goal, someone else may come to identify what our research project is about, and perhaps, publish it as their own. To avoid this, I must do two things: 1) Avoid detailing the particulars of what my research team and I are investigating before official publication 2) Avoid posting media of our specific procedure on a given specimen. I wish to share more with you, but unfortunately, until publication, I am limited to what EXACTLY I can say. Just know that alongside my lab protocols and techniques I perform, there's a lot more going on that I can't reveal right now. :)<br />
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One of the members of my lovely research team had her birthday this week-Andrea! Just because we are also working intensely to meet deadlines and submit papers doesn't mean we can't sit back and have a little bit of fun! To celebrate her birthday, one of the other members of the team, Cori, bought a delicious cake and brought it in. Every person on the research team, including Dr. McConnell, came in to celebrate and sing a version of "Happy Birthday" in their respective languages. After we celebrated, the inevitable discussion of what needs to be done in the lab and problems some of us are encountering in meeting particular deadlines ensued. Even though I have officially been a member of the lab for less than 3 weeks, I already feel like I've been here for years. It's definitely an enchanting feeling.<br />
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<tr><td class="tr-caption" style="text-align: center;">The Birthday Cake!</td></tr>
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<tr><td class="tr-caption" style="text-align: center;">Happy Birthday!</td></tr>
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<tr><td class="tr-caption" style="text-align: center;">The Research Team! (missing a few)</td></tr>
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Furthermore, I continued to make more TBST solution in the lab. TBST is probably one of the most commonly used solutions in our research lab because we are dealing with a high volume of Western blots. I also learned how to make a dilute solution of an antimicrobial to place in our cell incubator. It requires 5 mL of a blue substrate called AquaClean and 1000 mL (or 1 L) of distilled water. This may sound easy, but there has to be a lot of precision to how solutions are mixed and what type of "distilled water" is used. There's double distilled water, distilled water, and tap water in the lab. Some solutions require double distilled water (commonly labeled as ddH<sub>2</sub>O) while others require only distilled water (commonly abbreviated as DW). The pipetting procedure also is quite fastidious--one must spend a certain amount of time re-filling the pipette and emptying it out before all the substrate is removed. The solution must also be mixed thorougly in order to reach proper homogeneity.<br />
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjC4IXn9oHPNTMDY36kfRRjg_9dBebzB2FmMYm1gYxaosinPl0xeCqNab3sBYrBzxXJRuMRjlsYEDTkXWgks82J9AFNijuJ-1KpzOSiNybYUmU9McGz3WMX39ofzkFQRJvDV13QleNXP2Y/s1600/P6050029.JPG" imageanchor="1" style="clear: left; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" height="200" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjC4IXn9oHPNTMDY36kfRRjg_9dBebzB2FmMYm1gYxaosinPl0xeCqNab3sBYrBzxXJRuMRjlsYEDTkXWgks82J9AFNijuJ-1KpzOSiNybYUmU9McGz3WMX39ofzkFQRJvDV13QleNXP2Y/s200/P6050029.JPG" width="150" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">AquaClean Substrate</td></tr>
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<tr><td class="tr-caption" style="text-align: center;">Dilution</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEg6YqpjX9VIRuZRKhEqn2gUzW0D5wwxKfPSn_RqlCvHp27bF5xCXTSQbB0YMrqHdOTHXDeyYLNYM4KsCNadzKi09nyo5GiA7eRYU0q2t3IZuYSYerhIvYwrwj1t2eO-QwvaQdTZE0EKkfo/s1600/P6050031.JPG" imageanchor="1" style="clear: right; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEg6YqpjX9VIRuZRKhEqn2gUzW0D5wwxKfPSn_RqlCvHp27bF5xCXTSQbB0YMrqHdOTHXDeyYLNYM4KsCNadzKi09nyo5GiA7eRYU0q2t3IZuYSYerhIvYwrwj1t2eO-QwvaQdTZE0EKkfo/s200/P6050031.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Pan</td></tr>
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<tr><td class="tr-caption" style="text-align: center;">Cell Incubator</td></tr>
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With all of this TBST solution-making, it's not surprising that I also ran out of the 10x TBS, the substrate used to make the TBST solution. This was a rather lengthy and tedious process, especially since it requires the use of a pH meter and careful pipetting. First, I added 24.2 g of Tris base and 80 g of NaCl to a plastic beaker. Then, I stirred the contents after adding 800 mL of ddH<sub>2</sub>O . Because TBS is a buffer, I also had to ensure that the pH of was adjusted to a certain level--7.6 In order to do this, I had to standardize the pH electrode with 3 different pH level solutions (4, 7, and 10). I adjusted the pH of my solution by adding HCl to lower the pH and by adding NaOH to raise the pH until I reached 7.6 This took a while because one drop could really make a big difference in what the pH meter read, and I did not want to go over or under the level needed. Last, I added the remaining ddH<sub>2</sub>O to the solution to make 1 L total.<br />
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgDSWV1bts-h9z4n5JaGgQ7BXr5H0IzEMfkUtemrwxMihFQE_Ves95Z9t7Bp1ecfBKLQtQmdjfL3GlrNOtNmPlLhkbuv8FRxl4RwbAoQjsHSfklo3VDRMHNJJ240RnsC0Eh3nLAJ1M8ANs/s1600/P6060034.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="200" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgDSWV1bts-h9z4n5JaGgQ7BXr5H0IzEMfkUtemrwxMihFQE_Ves95Z9t7Bp1ecfBKLQtQmdjfL3GlrNOtNmPlLhkbuv8FRxl4RwbAoQjsHSfklo3VDRMHNJJ240RnsC0Eh3nLAJ1M8ANs/s200/P6060034.JPG" width="150" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Tris Base</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEh4ApXYVQHy5lLrmMQlz5QuECLHEd9if0KiELthYb9xMvaDoZ0ddkfiprGjHJ5QGZsJwN18xQntGMu5tZqnzFil-IjKROND2H7l79ZaTgzUQ10GQSbbXcsxQuRnHyZQIkHOiTnmemVmguc/s1600/P6060035.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="200" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEh4ApXYVQHy5lLrmMQlz5QuECLHEd9if0KiELthYb9xMvaDoZ0ddkfiprGjHJ5QGZsJwN18xQntGMu5tZqnzFil-IjKROND2H7l79ZaTgzUQ10GQSbbXcsxQuRnHyZQIkHOiTnmemVmguc/s200/P6060035.JPG" width="150" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Sodium Chloride</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEikQmM-6xPBYuhzuN4LA9NH-5l9z-3gM7pROlgmSgqxJCyCvP6IhEO_yCXjPYBovwGI8pHa8OFML3T_k_BmM8rRHmTJkbB_LqodPXSu2oRjs9QRgklb58DF-kyxevenuYuUrpSv5uLxPx4/s1600/P6060037.JPG" imageanchor="1" style="clear: right; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEikQmM-6xPBYuhzuN4LA9NH-5l9z-3gM7pROlgmSgqxJCyCvP6IhEO_yCXjPYBovwGI8pHa8OFML3T_k_BmM8rRHmTJkbB_LqodPXSu2oRjs9QRgklb58DF-kyxevenuYuUrpSv5uLxPx4/s200/P6060037.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">pH Standards</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjgzBWkLRidmDv4ERz59uhH0GSjzEoYe9Llw23bq8nTU834f-heKVsmtEiakpb5rtfAzJf-_gJKkGanrm7VCjW2el6tFOuUWlFYHgtof4vlkiDO1yB3MsCw6qcXptNf7AgBbBexIpUSljg/s1600/P6060038.JPG" imageanchor="1" style="clear: left; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjgzBWkLRidmDv4ERz59uhH0GSjzEoYe9Llw23bq8nTU834f-heKVsmtEiakpb5rtfAzJf-_gJKkGanrm7VCjW2el6tFOuUWlFYHgtof4vlkiDO1yB3MsCw6qcXptNf7AgBbBexIpUSljg/s200/P6060038.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Standardizing the pH meter</td></tr>
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<tr><td class="tr-caption" style="text-align: center;">Adding HCl and NaOH to adjust the pH reading</td></tr>
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<tr><td class="tr-caption" style="text-align: center;">Final pH-7.6</td></tr>
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<tr><td class="tr-caption" style="text-align: center;">The Final Product-10x TBS!</td></tr>
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This week was also very unique because I experienced my first team meeting! All of the research members and our PI, Dr. McConnell, assembled in a conference room to discuss the progress of our individual projects/experiments. Since there are many deadlines to meet, the tone of the conversation was tense and hasty. Dr. McConnell expressed the necessity to identify specific substrates in one of the members of my research team's experiment. This is extremely important because in order for a research paper to be published, it needs to answer a question with very specific results. If not, then the paper may not be of value to even publish. A general answer or a general piece of evidence is not sufficient enough for publication. Furthermore, we talked about what other experiments needed to be performed while also being mindful of the fact that these experiments must be replicated several times in order to yield a significant conclusion. Although my research team seemed a bit stressed out with what seemed to be a conversation of never-ending experiments and things to do, I promised to help out as much as I could. Dr. McConnell even reminded me to help out with several of the projects so that these looming deadlines could be met.<br />
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEg6pkPYJv9oPChWG_8PdTG4JCFCLlimk0gQhHFWaxylIOMhyIkFvYL8hyphenhyphenVelez25FhnhrFzs1syC-1slroge2qkuO94Vj7FJfpgHcLPHgSRaTEkuJ9aDNPido-cqWrMaRd5VbDT67Bx5DU/s1600/P6070046.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEg6pkPYJv9oPChWG_8PdTG4JCFCLlimk0gQhHFWaxylIOMhyIkFvYL8hyphenhyphenVelez25FhnhrFzs1syC-1slroge2qkuO94Vj7FJfpgHcLPHgSRaTEkuJ9aDNPido-cqWrMaRd5VbDT67Bx5DU/s200/P6070046.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Part of the Meeting </td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhwtHoha6IedUGvzQswCiN-1WnSErlz-dm3eZqQl5XyPpE5-C5u5EFm3RC33bAbEQ2Bdn0rOhy3QKNuTyjCHCcHgYVstDkL5LxCYBj-XFjm0izKE81N_PmTjiJNUxi9BucV7nxoucuJYn4/s1600/P6060045.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhwtHoha6IedUGvzQswCiN-1WnSErlz-dm3eZqQl5XyPpE5-C5u5EFm3RC33bAbEQ2Bdn0rOhy3QKNuTyjCHCcHgYVstDkL5LxCYBj-XFjm0izKE81N_PmTjiJNUxi9BucV7nxoucuJYn4/s200/P6060045.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">DNA Gel Electrophoresis </td></tr>
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<a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhuASnZFL7jw45XWhh0PjBvETJxX9kBVSIeb2s6YwB2KGkCLt5oovMZcqEh64chIe7_UnkOGdeMQYQL0eaDqBcLi9KPk_WDwFKAj0y_EQ3aOGWEGo98Z17kfl1cSXX97zxDB2RO6o118FE/s1600/P6080050.JPG" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhuASnZFL7jw45XWhh0PjBvETJxX9kBVSIeb2s6YwB2KGkCLt5oovMZcqEh64chIe7_UnkOGdeMQYQL0eaDqBcLi9KPk_WDwFKAj0y_EQ3aOGWEGo98Z17kfl1cSXX97zxDB2RO6o118FE/s200/P6080050.JPG" width="200" /></a><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEh6_7D-1EnDd_UPzGVMIA9ft6jZ4EC3yAJDW8msur-7vQW11GgYG8ldapWP7mSPP4uGI6I6s-tuK1CH1B1i96JFyEd5gJMyQuJIcSt3Yk4xv1rGexC2A8cF_KcBgNQF1etxQpqraRaNMLo/s1600/P6080053.JPG" imageanchor="1" style="margin-left: 1em; margin-right: 1em;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEh6_7D-1EnDd_UPzGVMIA9ft6jZ4EC3yAJDW8msur-7vQW11GgYG8ldapWP7mSPP4uGI6I6s-tuK1CH1B1i96JFyEd5gJMyQuJIcSt3Yk4xv1rGexC2A8cF_KcBgNQF1etxQpqraRaNMLo/s200/P6080053.JPG" width="200" /></a></div>
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<tr><td class="tr-caption" style="text-align: center;">Liquid Nitrogen (used to instantly freeze collected tissue for storage)</td></tr>
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<br /></div>Lauren Tolathttp://www.blogger.com/profile/05812297914953765766noreply@blogger.com0tag:blogger.com,1999:blog-8500808137876637614.post-46008170512881224752012-06-01T12:50:00.001-07:002012-06-25T20:29:08.483-07:00Week 2-Solutions and Dilutions<div class="separator" style="clear: both; text-align: center;">
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With the completion of my second week, I feel that I am slowly getting adjusted to the lifestyle and work involved in performing research. Many of the graduate students, as well as my fellow mentor, invest long hours into their individual experiments (which are all quite lengthy and cumbersome) in order to meet deadlines. Even though I was warned about the unfortunate "boring" and "slow" part of research, I am amazed at the progress I have been able to witness in the last two weeks. I am also extremely honored to be given the opportunity to work hand-in-hand with such knowledgeable and driven people during my undergraduate experience. I am, indeed, the youngest person to walk around in the lab, but I feel as if I am happily accepted into one big research family.<br />
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For the second week, I continued to wash the Western Blots with the TBST solution and saturate them in a milk+antibodies solution. Afterwards, I participated in making an ECL solution. ECL stands for enhanced chemiluminescence and it allows detection of minute quantities of a biomolecule. This occurs because the reaction formed by the mixture emits light when it decays to a singlet carbonyl. Afterwards, we took the Western Blots to a dark room and developed the protein bands on a film. The protein bands are in correspondence to the antibodies that they are saturated with (i.e.-GAPDH).<br />
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiWOaeFtld3oJMpvCHiSPz9aJ6AhdD3mVjtfLtI-lG0tjpcCjjOp1Z8BE47aU_whleZnCOUJjXGDRqNItJGVDV7CUN8fKQH4efPjRXJTrcbtxxhVnJ61CaY3hI2HFQ6MTpWlJh0gEwaBaw/s1600/P5290002.JPG" imageanchor="1" style="clear: left; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiWOaeFtld3oJMpvCHiSPz9aJ6AhdD3mVjtfLtI-lG0tjpcCjjOp1Z8BE47aU_whleZnCOUJjXGDRqNItJGVDV7CUN8fKQH4efPjRXJTrcbtxxhVnJ61CaY3hI2HFQ6MTpWlJh0gEwaBaw/s200/P5290002.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Substrate used to make ECL</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhWmYgGoWcVTG6LdydiAhZzs0gWg8fd5_IZzYRvsemnTlJz-WOlKZtUZ_3JDNEvb3tuQpkLmlwRmvkQ71Dfqu1TWH6nnnDsf35ZAtj5i0oDgdw4tYM1an3lwvVjg0kRVcx5Oo7r34tNz2o/s1600/P5290003.JPG" imageanchor="1" style="clear: right; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhWmYgGoWcVTG6LdydiAhZzs0gWg8fd5_IZzYRvsemnTlJz-WOlKZtUZ_3JDNEvb3tuQpkLmlwRmvkQ71Dfqu1TWH6nnnDsf35ZAtj5i0oDgdw4tYM1an3lwvVjg0kRVcx5Oo7r34tNz2o/s200/P5290003.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Another set of substrates used to make ECL</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgXc5efDW0AyMDI4l_KObVEzlJOsGnYBI7Sn2U2Zasfya6Gibamy-n4XTzd75s-vQr28-W6a3NHr4lZ9sFdlrsOxprJ52RlnyWhjh56UOnkGxCT9gd-q2L40NU0AmsywXCKYf8V4IV2xBI/s1600/P5290004.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgXc5efDW0AyMDI4l_KObVEzlJOsGnYBI7Sn2U2Zasfya6Gibamy-n4XTzd75s-vQr28-W6a3NHr4lZ9sFdlrsOxprJ52RlnyWhjh56UOnkGxCT9gd-q2L40NU0AmsywXCKYf8V4IV2xBI/s200/P5290004.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Substrate Bottles</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgFWd3m1r8AEtIybhYuomTWKQ2j4UZPWZGBWaMhF2tRj7CDyS3hSDQPeYrW7x1H5xFyoDgcUZ7_NPVgkacxf2UGzSpmtuoa9O13sTshpg5LPvN_oTn-vu6cVQfoB6yU361LqLdvf_pdmgM/s1600/P5290005.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="200" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEgFWd3m1r8AEtIybhYuomTWKQ2j4UZPWZGBWaMhF2tRj7CDyS3hSDQPeYrW7x1H5xFyoDgcUZ7_NPVgkacxf2UGzSpmtuoa9O13sTshpg5LPvN_oTn-vu6cVQfoB6yU361LqLdvf_pdmgM/s200/P5290005.JPG" width="150" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Developed Western Blot</td></tr>
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I also prepared more TBST solution. This requires a 10x dilution (100 mL of TBST, 900 mL of double distilled water, and 1 mL of a detergent based liquid called Tween). I also prepared more ECL solution and more of a milk+antibodies solution. I also washed the Western Blots (after it had been taken to the dark room to be processed) with TBST two times (5 minutes each) and then a stripping buffer (8 mL for the large box and 5 mL for the two smaller boxes) for 12 minutes. Afterwards, I washed the boxes with a TBST solution two times for 5 minutes each. Last, I added milk+antibodies for 1 hour.<br />
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiEmlW4xS3d07NF87nL0YGl-OsUFl_uww-rEPsPyy9EmeIQUD7Bne-6OlIe_6g-2ZDu7FvGQqsqOVhrQm-l8-T9KTL7QXl5fzAOVUMrlzHtO89lB8chtJJacK71o9_m4PQgH0kEhuv3X8I/s1600/P6010014.JPG" imageanchor="1" style="clear: left; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEiEmlW4xS3d07NF87nL0YGl-OsUFl_uww-rEPsPyy9EmeIQUD7Bne-6OlIe_6g-2ZDu7FvGQqsqOVhrQm-l8-T9KTL7QXl5fzAOVUMrlzHtO89lB8chtJJacK71o9_m4PQgH0kEhuv3X8I/s200/P6010014.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Making TBST-diluting 10x</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhd6_zAlQMK-a1r8Du-bXHBS-9E9ZDAl6C8uovOzMG1r-ISwcfehrYvynLVYeZeBllbCQcv20pY3Xl-zTNE2ATdkzEdZsGUzB8Kawq6eA6vaJj4EEPku6lDFafcuA2kmLg2mGi-r9QiL8M/s1600/P6010015.JPG" imageanchor="1" style="font-size: 12px; line-height: 17px; margin-left: auto; margin-right: auto;"><span style="color: #404040; font-family: 'Lucida Grande', Verdana, Arial, sans-serif;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhd6_zAlQMK-a1r8Du-bXHBS-9E9ZDAl6C8uovOzMG1r-ISwcfehrYvynLVYeZeBllbCQcv20pY3Xl-zTNE2ATdkzEdZsGUzB8Kawq6eA6vaJj4EEPku6lDFafcuA2kmLg2mGi-r9QiL8M/s200/P6010015.JPG" width="200" /></span></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;"><span style="font-family: Times, 'Times New Roman', serif; font-size: x-small;"><span style="line-height: 17px;">Making TBST-adding 900 mL of dd</span><span style="background-color: white; line-height: 17px; text-align: justify;">H</span><sub style="background-color: white; line-height: 17px; text-align: justify;">2</sub><span style="background-color: white; line-height: 17px; text-align: justify;">O</span></span></td></tr>
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<tr><td class="tr-caption" style="text-align: center;">Making TBST-adding 1 mL of Tween</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjPp5KG3MVNNK0benm8o6GqCVnX4oUzkyd2J8JI7bGPr5BXch_DiDjL2snq8iAXL-xGOUMXLpgtWmoB7D3cToNp0I0st35rT8lYL1ADj3KHrNNFhyphenhyphenDz5ONiWIsudKqLXDFz1aijn2R6xsQ/s1600/P6010020.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="200" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjPp5KG3MVNNK0benm8o6GqCVnX4oUzkyd2J8JI7bGPr5BXch_DiDjL2snq8iAXL-xGOUMXLpgtWmoB7D3cToNp0I0st35rT8lYL1ADj3KHrNNFhyphenhyphenDz5ONiWIsudKqLXDFz1aijn2R6xsQ/s200/P6010020.JPG" width="150" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Making TBST-Final product with an updated label</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEi4suKmX4IHs-UA10DdR_Ul1vNdEaCmFUPH249xtn297J5M0LnjQ7iZg9yNeCwcmQETBO6VKDI9WDuag5VyKXvlGvEiPFCP3zp6Xcm_-mFLMrpFGp9ybXFKOTW7kvMvqjJQnfOKKPTtdRU/s1600/P5290006.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><span style="font-size: x-small;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEi4suKmX4IHs-UA10DdR_Ul1vNdEaCmFUPH249xtn297J5M0LnjQ7iZg9yNeCwcmQETBO6VKDI9WDuag5VyKXvlGvEiPFCP3zp6Xcm_-mFLMrpFGp9ybXFKOTW7kvMvqjJQnfOKKPTtdRU/s200/P5290006.JPG" width="200" /></span></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Making the Milk Solution </td></tr>
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<tr><td class="tr-caption" style="text-align: center;">Making TBST-Stirring all contents with this device</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjdmzQxWyMapZF-cOqu-psndR8a4HyZAscB5Dg9s7nclecinw5VDjLr6B2PQLF3_IFwn-FxjjIelIv2vKmwtxnZwfY07KTDgKO_rzZ_DcPS3nIwSNAG2VJ1tO-qxk1V2RvbsnmRxQbrNFk/s1600/P5300008.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="200" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjdmzQxWyMapZF-cOqu-psndR8a4HyZAscB5Dg9s7nclecinw5VDjLr6B2PQLF3_IFwn-FxjjIelIv2vKmwtxnZwfY07KTDgKO_rzZ_DcPS3nIwSNAG2VJ1tO-qxk1V2RvbsnmRxQbrNFk/s200/P5300008.JPG" width="150" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Stripping Buffer used in Western Blots</td></tr>
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<br /></div>Lauren Tolathttp://www.blogger.com/profile/05812297914953765766noreply@blogger.com1tag:blogger.com,1999:blog-8500808137876637614.post-13526609807316458002012-05-25T14:07:00.001-07:002012-06-25T20:30:02.049-07:00Week 1-Protein Assays and an Assortment of Duties<div class="separator" style="clear: both; text-align: center;">
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As the end of my first week approaches, I can already tell that research will be one of the best moments of my undergraduate experience. The environment, my professor, and my research team are all phenomenal and I am so thrilled to be a part of this. <br />
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On the first day of lab, I was oriented to the progress in the overall research project that is currently going on.<br />
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhpBarQkeTahIfiPi1KTo3WccqDqqzF1-QnGPsODqr2t6cW1oEQEn7ZefljJF_CHesiFnAvPiPt_R0bQUUuo8ihrXEgFZNKC1n4fWuN6T69T4dui2b1VUVfDgvulCmaZhZ7rNBmzuCBHeg/s1600/P5200047.JPG" imageanchor="1" style="clear: left; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhpBarQkeTahIfiPi1KTo3WccqDqqzF1-QnGPsODqr2t6cW1oEQEn7ZefljJF_CHesiFnAvPiPt_R0bQUUuo8ihrXEgFZNKC1n4fWuN6T69T4dui2b1VUVfDgvulCmaZhZ7rNBmzuCBHeg/s200/P5200047.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="font-size: 13px; text-align: center;">My Lab Setting<br />
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<span style="text-align: left;">My research activities included making dilutions from more concentrated solutions (or stock solutions) and washing collected tissues with a PBS (saline) solution. I had to be very careful in this procedure because I know my graduate student mentor worked tediously to isolate and collect such delicate tissue. I also participated in making more of a PBS solution and making a dilute solution of the buffer, TBST. </span><br />
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjiePbBkjlYKPefH2McRayIG4t3LiMvP1Ze5VJcalIw4j3lKbVl3jTGBwpSJgQaVcpvV1y4QmaVfVlwl_wvpOr68loU63dXBcVyBzgJSAqYIco2yPKayVab5GctC4iVGym7oGl8q-j7nfs/s1600/P5210003.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjiePbBkjlYKPefH2McRayIG4t3LiMvP1Ze5VJcalIw4j3lKbVl3jTGBwpSJgQaVcpvV1y4QmaVfVlwl_wvpOr68loU63dXBcVyBzgJSAqYIco2yPKayVab5GctC4iVGym7oGl8q-j7nfs/s200/P5210003.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Washing Collected Tissue with PBS Solution</td></tr>
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I also performed a protein concentration measurement with a BCA protein assay kit. Protein assays are performed to measure the total level of protein in a solution. A series of dilutions of known concentration are prepared from the protein and assayed alongside the unknowns before the concentration of each unknown is determined based on the standard curve. The first step in this process is to prepare our diluted albumin standards, known as the BSA. These will standardize our results during the spectrophotometer readings. This entire process is quite lengthy and it requires extremely careful use of several different types of pipettes to insert a precise amount of solute (as low as 2 microliters). The second step in this process is to prepare the BCA Working Reagent which will be used to indicate the concentration of a particular protein in our series of dilutions. The Reagent changes color (green or purple) depending on the concentration of the protein present in the solution. The Reagent is then added to the protein solutions. The third step involves incubating the protein solutions at 37 degrees Celsius for 30 minutes. After incubation, I used a spectrophotometer to measure the absorbance of all the samples. This procedure had to be done very quickly (within 10 minutes) due to temperature-sensitive color development. I finally recorded all the data from my spectrophotometer on an Excel spreadsheet that included averaged values and the actual concentrations (the spectrophotometer reading multiplied by 25 because we performed a 25x dilution to prepare the solution at the beginning). The BSA standards that I prepared are stored in the freezer for later use. Overall, this protein assay matched and confirmed the data values that had previously been collected.<br />
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<tr><td class="tr-caption" style="text-align: center;">Results Printed from the Spectrophotometer</td></tr>
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<tr><td class="tr-caption" style="text-align: center;">Protein Samples Stored at 4<span style="background-color: white; color: #222222; font-family: arial, sans-serif; font-size: x-small; line-height: 16px;">°</span>C</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhPmx9IutY6BuPH4ITrmuSZSkHP7XSys7Xp0kZN1cnBPjADSyFruqRjxnjp1qZpQZcBNq7qDnKVfoPwNZyx62nGZGe1JPiGL7KQTE9nRryiWs1zOV2yAkqgkDliTk0SQDf2GXkatrbaguM/s1600/P5230011.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEhPmx9IutY6BuPH4ITrmuSZSkHP7XSys7Xp0kZN1cnBPjADSyFruqRjxnjp1qZpQZcBNq7qDnKVfoPwNZyx62nGZGe1JPiGL7KQTE9nRryiWs1zOV2yAkqgkDliTk0SQDf2GXkatrbaguM/s200/P5230011.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Eppendorf tubes with Standards and Samples</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjdSAvM5B0ZB3hq34GKvkG6GsRnHpwX-i-7YXWepARGseTp7OpIwDIP216smM-7bugEXAOvgTJa5vcz8krMZpx1uL5wM_p5e5oYHCel1lhR6UxoOAkh9hV-znJqYjI7l7BmLBvA0UNUkL0/s1600/P5220007.JPG" imageanchor="1" style="clear: left; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" height="200" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjdSAvM5B0ZB3hq34GKvkG6GsRnHpwX-i-7YXWepARGseTp7OpIwDIP216smM-7bugEXAOvgTJa5vcz8krMZpx1uL5wM_p5e5oYHCel1lhR6UxoOAkh9hV-znJqYjI7l7BmLBvA0UNUkL0/s200/P5220007.JPG" width="150" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Pipetting </td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjd7MZ3D0AJVQsMXVueM7EqFLdzI09MCg-udb2W505x69qJ02dCIe1fqMg8S0k1dAWdkVE4VCeahj-CZrIdEn0qRLBpxdopfEegh6K6UUgG9lYnLGRoTzjsIspNOwVslcs_5TPMBr-fC-E/s1600/P5220009.JPG" imageanchor="1" style="clear: left; margin-bottom: 1em; margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEjd7MZ3D0AJVQsMXVueM7EqFLdzI09MCg-udb2W505x69qJ02dCIe1fqMg8S0k1dAWdkVE4VCeahj-CZrIdEn0qRLBpxdopfEegh6K6UUgG9lYnLGRoTzjsIspNOwVslcs_5TPMBr-fC-E/s200/P5220009.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">Protein Assay Kit</td></tr>
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<tr><td style="text-align: center;"><a href="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEj3Pw_QqcsII4GgqMqMJCwEgEwk4Gx6MmybbFpTA_0M9_IFCMzxBmO2jaciaK3nHDgWIiova-9_sSOFMm9J7TcLDPF0LxxuM8GtjIsGlZN_SD-ccFA0ciU4F9_WoIZ5TGKP4Hyxe5jsFbI/s1600/P5230014.JPG" imageanchor="1" style="margin-left: auto; margin-right: auto;"><img border="0" height="150" src="https://blogger.googleusercontent.com/img/b/R29vZ2xl/AVvXsEj3Pw_QqcsII4GgqMqMJCwEgEwk4Gx6MmybbFpTA_0M9_IFCMzxBmO2jaciaK3nHDgWIiova-9_sSOFMm9J7TcLDPF0LxxuM8GtjIsGlZN_SD-ccFA0ciU4F9_WoIZ5TGKP4Hyxe5jsFbI/s200/P5230014.JPG" width="200" /></a></td></tr>
<tr><td class="tr-caption" style="text-align: center;">The Working Reagent</td></tr>
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On the fourth day, my primary focus was transferring the membranes of previously completed Western </div>
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blots into sealed plastic bags. This involved taking the membrane out of its container, transferring it to a plastic bag, adding 0.1% TBST (a buffer) to the plastic bag, and then sealing it with a machine. I cleaned out the black fluorescence boxes that previously contained the Western blots and shelved them so that they can be used for the Western blots that need to be completed in the near future.</div>
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<tr><td class="tr-caption" style="text-align: center;">Transferred Membranes from Western Blots</td></tr>
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On the last day of my first week, I assisted in finishing some of the Western blots. I emptied out the solution in the black boxes that had already been incubated for a certain time, incubated the Western blots with a stripping buffer for 8 minutes two times, then proceeded to wash the membranes with a TBST solution two times for 10 minutes each. For each step, I placed the membranes on a rocker in order to keep the membrane saturated throughout the incubation period. After this, I added a milk solution (1.5 g of Nonfat Dry Milk with 30 mL of TBST) and incubated the membranes in this for an hour. Once the hour was completed, I saturated the membranes in a milk solution+antibodies for 2 hours. After the 2 hours was completed, I washed the Western blots with the TBST solution 3 times for 5 minutes each. Once washing was complete, I incubated the membranes with another milk solution+antibodies for 1 hour.<br />
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<tr><td class="tr-caption" style="text-align: center;">"Rocker" Machine used to Keep Western Blots Saturated </td></tr>
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<tr><td class="tr-caption" style="text-align: center;">Milk+Antibodies used in Western Blotting</td></tr>
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That's it for now! See you at the end of next week!</div>
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~Lauren Tolat</div>
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</div>Lauren Tolathttp://www.blogger.com/profile/05812297914953765766noreply@blogger.com1tag:blogger.com,1999:blog-8500808137876637614.post-27929330979412994062012-05-15T12:49:00.001-07:002012-08-09T10:04:51.740-07:00Introduction to My Blog and Summer Research Plans<br />
Hello all!<br />
<br />
My name is <a href="http://www.uh.edu/tieronescholars/meet-our-scholars/2011-scholars/tolat-lauren.php">Lauren Tolat</a> and I have just completed my first year at the University of Houston. I am a Tier One Scholar, a Pre-Pharmacy major, and a member of the Honors College. This summer, I will be embarking on an exciting research opportunity with the College of Pharmacy, my faculty advisor, <a href="http://www.uh.edu/pharmacy/directory-home/pps-faculty/bradley-mcconnell/index.php">Dr. Bradley K. McConnell</a>, and my graduate student mentor, Qiying Fan. I will be sharing my weekly logs, updates, and news with all of you in regards to this slow yet beautiful thing called research! <span style="text-align: left; text-indent: -24px;">Thanks to my Tier One scholarship and the help of friendly UH faculty members, I am privileged to have this opportunity. </span><br />
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I will be working in the research lab from May 21st through August 10th. Here's a summary of what I will be working on:<br />
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<span style="font-family: inherit;"><b><u>Project title</u></b><b>:</b></span></div>
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<span style="font-family: inherit;">Cell Signaling in Health and in Cardiovascular Disease</span></div>
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<span style="font-family: inherit;"><b><u>Objectives</u></b><b>:</b></span></div>
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<span style="font-family: inherit;">Heart disease is the number one cause of death in the
USA, affecting more than 12 million Americans. Furthermore, nearly 5 million
patients in the USA have heart failure; where the heart is unable to
efficiently pump blood. Research interests in the laboratory focuses on the
genetics and physiology of heart disease and atherosclerosis, as it relates to
normal healthy and diseased hearts and vasculature. In particular, our goals
are to define mechanisms and to investigate strategies to improve cardiac and
vascular smooth muscle (VSMC) function in cardiovascular disease.</span></div>
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<span style="font-family: inherit;">There are several projects
in Dr. McConnell’s laboratory focusing on
targeted disruption of beta-adrenergic signaling to alter cardiac
contractility. In particular, the lab is
focused on studying the signal transduction
pathways in the heart .This course will give the
student, Lauren Tolat, the opportunities to obtain research
experience and learn advanced laboratory techniques. </span></div>
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<u style="font-family: inherit; font-weight: bold;">Lab Techniques</u><span style="font-family: inherit;">:</span></div>
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<span style="font-family: inherit;">1. Cardiomyocyte and VSMC
isolation<o:p></o:p></span></div>
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<span style="font-family: inherit;">2. Histology and
Immunohistochemisty (sectioning, staining, and imaging)<o:p></o:p></span></div>
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<span style="font-family: inherit;">3. SDS-PAGE gel to separate
proteins based on molecular weight <o:p></o:p></span></div>
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<span style="font-family: inherit;">4. Western blotting for total
protein and phospho-protein detection<o:p></o:p></span></div>
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<span style="font-family: inherit;">5. Data analysis, chart and
figure creation and presentation </span></div>
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<span style="font-family: inherit; text-align: left; text-indent: -0.25in;">Thanks for tuning in. I will have more to post in the forthcoming weeks.</span></div>
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<span style="font-family: inherit;">~Lauren Tolat</span></div>
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<br />Lauren Tolathttp://www.blogger.com/profile/05812297914953765766noreply@blogger.com0