User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/08/21: 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;">Lysate SDS-PAGE, qPCR AKAP5, Western blot AKAP79 and Gravin</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== | ||
* SDS-PAGE Lysate samples | * SDS-PAGE Lysate samples | ||
** Forgot antioxidant, added after 1h 11 min.; power failure @ 2h 40 min. → continued to blot (2h, 30V) | |||
** Ponceau staining does not look very nice compared to others | |||
** Gel is coomassie stained | |||
{| border="1" | {| border="1" | ||
|+'''4-12% Gradient NuPage Gel loading LYSATES hTERT D9 + HEK293, HeLa -S & Jurkat see codes [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/21|21June2010]]''' | |+'''4-12% Gradient NuPage Gel loading LYSATES hTERT D9 + HEK293, HeLa -S & Jurkat see codes [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/21|21June2010]]''' | ||
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! align="center" style="background:#f0f0f0;"|<u>10</u> | ! align="center" style="background:#f0f0f0;"|<u>10</u> | ||
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!Gel<br> | !Gel<br>Lysates | ||
| align="center"|[[Media:HiMark_protein_marker_-_Invitrogen.jpg|Invitrogen Marker]]<br> (8 μL) | | align="center"|[[Media:HiMark_protein_marker_-_Invitrogen.jpg|Invitrogen Marker]]<br> (8 μL) | ||
| align="center"|Jurkat <br>(20 μL) | | align="center"|Jurkat <br>(20 μL) | ||
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* Prepared cDNA | * Prepared cDNA | ||
* qPCR AKAP5 and GAPDH | * qPCR AKAP5 and GAPDH | ||
** Not enough master mix for HEK293 and Jurkat controls | |||
[[Image:AKAP79_expression.jpg|500px]] | [[Image:AKAP79_expression.jpg|500px]] | ||
* Western blot 2<sup>nd</sup> Antibody (α-Rabbit) 1:10.000 | * Western blot 2<sup>nd</sup> Antibody (α-Rabbit) 1:10.000 | ||
** Blot #1 [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/16|16August2010]], Loading control was already performed [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/19|19August2010]] | |||
[[Image:21082010_Lysates_Immunoblot_AKAP250_and_79.tif|500px]] | |||
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==Materials & Methods== | ==Materials & Methods== |
Revision as of 06:49, 23 August 2010
<html><style type="text/css"> .todo { color: red } .done { color: green} </style></html>
Lysate SDS-PAGE, qPCR AKAP5, Western blot AKAP79 and Gravin | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | ||||||||||||||||||||||
Summary
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