User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/08/21

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Lysate SDS-PAGE, qPCR AKAP5, Western blot AKAP79 and Gravin Main project page
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Summary

  • 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
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
Lysates 		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)
  • Prepared cDNA
  • qPCR AKAP5 and GAPDH
    • Not enough master mix for HEK293 and Jurkat controls

  • Western blot 2nd Antibody (α-Rabbit) 1:10.000


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