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

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Summary

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

Notes

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

Results

  • Immunoblot

24062010 AKAP immunoblot.png

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

Discussion

  • 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