User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/08/10

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Summary

  • 4.450 mg st-Ht31 was diluted in 159 µL DMSO
  • ELISA
    • NOTE: Plate #1 well C1 received 200 µL of S0 instead of CSE, well D1 received 400 µL and also did not receive CSE. Endvolumes were still 600 µL
1.100810 ' S0+DMSO S0+DMSO S0+DMSO st-Ht31 (50 µM) st-Ht31 (50 µM) st-Ht31 (50 µM)
1 2 3 4 5 6
CTR A
CSE (15%) B
CSE+db-cAMP (500 µM) C
CSE+ mPKI (400 nM) D
02.100810 ' Control 8Pcpt 6Bnz Ht31+6Bnz DMSO Ht31
1 2 3 4 5 6
CTR A
CTR B
CSE C
CSE D
  • SDS-PAGE
4-12% Gradient NuPage Gel loading hTERT D9 + HEK293, HeLa -S & Jurkat see codes 21June2010
# 1 2 3 4 5 6 7 8 9 10 11 12
Gel
cAMP prec.
Invitrogen Marker
(8 μL)
Jurkat (LYSATE)
(20 μL)
HeLa -S
(20 μL)
HEK293
(20 μL)
Biorad Marker
(8 μL)
C1
(20 μL)
C2
(20 μL)
C3
(20 μL)
S1
(20 μL)
S2
(20 μL)
S3
(20 μL)
X

Results

  • Ponceau staining cAMP precipitation blot (4-12% NuPage)

Discussion

  • DONE Ow..