User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/05/31: Difference between revisions
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====Splitting cells==== | ====Splitting cells==== | ||
* Splitting cells into 12-wells? | * <del>Splitting cells into 12-wells?</del> | ||
*D9 to passage 25 | *D9 to passage 25 | ||
====Immunoblot==== | ====<del>Immunoblot</del>==== | ||
*Block membrane in 1% BSA TBST solution for >1 h @ RT | *Block membrane in 1% BSA TBST solution for >1 h @ RT | ||
*Incubate with primary antibody 2 h @ RT or ON @ 4 °C | *Incubate with primary antibody 2 h @ RT or ON @ 4 °C | ||
Line 79: | Line 79: | ||
**1:10.000 dilution in blocking solution | **1:10.000 dilution in blocking solution | ||
*Wash 3x 10 min. with blocking solution | *Wash 3x 10 min. with blocking solution | ||
====Preparing film==== | ====<del>Preparing film</del>==== | ||
* 1:1 ratio [[wikipedia:Chemiluminescence#Enhanced_chemiluminescence|ECL]] solutions (2 mL per membrane) | * 1:1 ratio [[wikipedia:Chemiluminescence#Enhanced_chemiluminescence|ECL]] solutions (2 mL per membrane) | ||
* Incubate membrane in ECL solution 5 min, RT (both sides of membrane) | * Incubate membrane in ECL solution 5 min, RT (both sides of membrane) |
Revision as of 09:11, 31 May 2010
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SDS-PAGE for AKAP79 immunoblot & RII overlay (HEK293) | <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> | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
SummarySDS-PAGE
Splitting cells
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