User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/10/11

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Summary

  • Blocking membrane of 08October2010 → Incubating membrane with RIIα (sc-909) and Gravin (animal 122)dilution 1:500 ON @ 4°C
  • Preparation of 2 10% Tris-glycine acrylamide gels
  • Dilution of samples, corrected to lowest concentration
Sample μL Sample μL dH2O
HEK293 (12/08/2010)61,588,5
HeLa -S (p17 (20.6.2010)26,0124,0
S0 (7.5.2010)236,90,0
S152,197,9
S2150,00,0
S353,896,2
C0 (7.5.2010)2470,40,0
C159,690,4
C236,1113,9
C3106,343,7
10% SDS-PAGE Gel loading hTERT D9 + Jurkat, HEK293 & HeLa -S see codes 21June2010 S0 & C0 are of 7.5.2010
# 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
Gel
Lysates
X Fermantas high range Marker
(5 μL)
HEK293
(30 μL)
HeLa -S
(30 μL)
Jurkat
(30 μL)
Fermantas high range Marker
(10 μL)
S0
(30 μL)
S1
(30 μL)
S2
(30 μL)
S3
(30 μL)
C0
(30 μL)
C1
(30 μL)
C2
(30 μL)
C3
(30 μL)
X
  • Run at 200 V
  • 10% gel was cut in 4 for
    • GAPDH (1:2000, mouse (<40kDa))
    • RIIα (1:1000, rabbit (40 - 75kDa))
    • AKAP79 (1:1000, rabbit? (75 - 130kDa)) (Santa Cruz)
    • AKAP250 (1:1000, Goat, (>130kDA)) (Santa Cruz)


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