User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/07/09: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Gravin Co-IP, cAMP precipation gel</span> | ||
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==Summary== | ==Summary== | ||
* AKAP250 immunoprecipitation | * AKAP250 immunoprecipitation | ||
* Gradient (4-12%) SDS-PAGE (NuPage) | * Gradient (4-12%) SDS-PAGE (NuPage) for cAMP Precipitations | ||
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|+'''Gel loading hTERT D9 + HEK293 & HeLa -S see codes [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/21|21June2010]]''' | |+'''Gel loading hTERT D9 + HEK293 & HeLa -S see codes [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/21|21June2010]]''' | ||
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==Notes== | ==Notes== | ||
- | * HeLa cells had less sample put too gel, both HEK293 and HeLa are very slimy | ||
* Forgot to lyse the ordered HEK293 cells | |||
* Blot was blocked ON in 1% BSA in TBS-T | |||
* Gel was Coomassie stained ON | |||
==Results== | ==Results== | ||
*Overnight autoradiograph of RII overlay did not change anything of the image of yesterday | *Overnight autoradiograph of RII overlay did not change anything of the image of yesterday | ||
*Ponceau of cAMP pull down SDS-PAGE | |||
[[Image:09072010_Ponceau.png|500px]][[Image:09072010_Coomassie.png|500px]] | |||
==Discussion== | ==Discussion== | ||
* | * Ponceau shows bands at higher altitude, maybe look for AKAP-Lbc next to AKAP250? | ||
* Anoying that both markers are not running equal on every height, but HiMark at least has nice sharp band @ 160 kDa on NuPage gels. Although transfer was less perfect | |||
* Coomassie staining shows not a 100% efficiency of transfer, and it shows bands in the control lanes that were not visible on Ponceau staining. Hopefully something did transfer | |||
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==Related== | ==Related== | ||
===Related entries=== | ===Related entries=== | ||
Revision as of 05:33, 29 July 2010
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 Gravin Co-IP, cAMP precipation gel
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Summary
Notes
Results
Discussion
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