User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/06/24: 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;">Immunoblot repeat</span> | ||
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html> </html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}} | |style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html> </html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}} | ||
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==Summary== | ==Summary== | ||
* | * To check if really nothing could be detected the same antibodies were used ON, repeat of [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/23|yesterday]]. | ||
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==Materials & Methods== | ==Materials & Methods== | ||
===Materials=== | ===Materials=== | ||
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==Notes== | ==Notes== | ||
* The rotating device to turn gels ON had broken down, but were switched to different machine by Philipp | |||
==Results== | ==Results== | ||
*{ | *Immunoblot | ||
[[Image:24062010_AKAP_immunoblot.png|500px]] | |||
{| border="1" | |||
|+'''Gel loading hTERT D9 + HEK293 see codes [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/21|21June2010]]''' | |||
! align="center" style="background:#f0f0f0;"|# | |||
! align="center" style="background:#f0f0f0;"|<u>1</u> | |||
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!Gel<br>cAMP prec. | |||
| align="center"|X | |||
| align="center"|HEK293<br>(20 μL) | |||
| align="center"|[[Media:HiMark_protein_marker_-_Invitrogen.jpg|Marker]]<br> (8 μL) | |||
| align="center"|C1<br>(20 μL) | |||
| align="center"|C2<br>(20 μL) | |||
| align="center"|C3<br>(20 μL) | |||
| align="center"|S1<br>(20 μL) | |||
| align="center"|S2<br>(20 μL) | |||
| align="center"|S3<br>(20 μL) | |||
| align="center"|X | |||
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==Discussion== | ==Discussion== | ||
* | * RIIα detection went fine now, no explanation why it didn't work before like the rest | ||
* The other antibodies still could not find anything | |||
** To see if it is due to the transfer to the membrane a new gel will be run and stained (Coomassie or Silver) | |||
** Another possibility is to do the RII overlay on the remaining blot of [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/22|22June2010]] to see if there is anything that was precipitated together with RII subunits | |||
* As a positive control for AKAP95 and AKAP450 [[wikipedia:HeLa|HeLa cells]] have been requested | |||
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Revision as of 01:50, 25 June 2010
<html><style type="text/css"> .todo { color: red } .done { color: green} </style></html>
 Immunoblot repeat
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Summary
Notes
Results
Discussion
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