User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/08/17: Difference between revisions

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* Blots blocked yesterday were treated with α-RIIα, α-AKAP95 (mouse) and α-AKAP450 to see what we can find in lysates despite the fact that transfer was not complete
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==Materials & Methods==
==Materials & Methods==

Revision as of 07:35, 17 August 2010

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Summary

  • According to the NuPage blotting manual adding 0.01% SDS might improve transfer to the membrane
    • Volume of blotting chamber is ~200 mL so adding 100 μL of 20% SDS solution should be fine
  • Blot of 16August2010 is repeated (only one gel)
    • NOTE: Only 10 µL for HEK293 cells and de Untreated samples a smaller amount of sample left (about 70-80% of CSE treated)
4-12% Gradient NuPage Gel loading LYSATES hTERT D9 + HEK293, HeLa -S & Jurkat see codes 21June2010
# 1 2 3 4 5 6 7 8 9 10
Gel
cAMP prec. 		Invitrogen Marker
(8 μL) 		Jurkat
(20 μL) 		HEK293
(10 μL) 		Biorad Marker
(8 μL) 		C1
(20 μL) 		C2
(20 μL) 		C3
(20 μL) 		S1
(<20 μL) 		S2
(<20 μL) 		S3
(<20 μL)
  • Blots blocked yesterday were treated with α-RIIα, α-AKAP95 (mouse) and α-AKAP450 to see what we can find in lysates despite the fact that transfer was not complete


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