User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/05/09

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Materials & Methods

Method

  • Put samples to 8% polyacrylamide gell
# 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
Sample Marker CSE beads CSE NAC CSE Lysate CSE (-) beads CSE (-) NAC CSE (-) Lysate x S0 beads S0 NAC S0 Lysate S0 (-) beads S0 (-) NAC S0 (-) Lysate marker

Notes

  • Slot 8 was filled and slot 15 had no marker added

Results

  • Ponceau staining of cAMP precipitation on 8% Gel

File:100509 ponceau cAMP & lysates.tif

Discussion

  • A band around 55 kDa is shown only concentrated in the cAMP pull down
    • This would be RII subunits showing that cAMP precipitation is succesful
  • cAMP treated beads pull down don't show any bands, indicating that cAMP prevented the binding of samples to cAMP bound to the beads