User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/06/24

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Immunoblot repeat

Summary

To check if really nothing could be detected the same antibodies were used ON, repeat of yesterday.

The rotating device to turn gels ON had broken down, but were switched to different machine by Philipp

**Gel loading hTERT D9 + HEK293 see codes 21June2010**
#	1	2	3	4	5	6	7	8	9	10
Gel cAMP prec.	X	HEK293 (20 μL)	Marker (8 μL)	C1 (20 μL)	C2 (20 μL)	C3 (20 μL)	S1 (20 μL)	S2 (20 μL)	S3 (20 μL)	X

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 22June2010 to see if there is anything that was precipitated together with RII subunits
As a positive control for AKAP95 and AKAP450 HeLa cells have been requested