Summary
cAMP precipitation
- Beads were washed as described 09April2010
- 50 μL of NuPage LDS sample buffer + DTT was added to the beads (800 μL LDS sample buffer + 200 μL 1 M DTT)
- Beads were put to 70 °C for 10 min.
- Samples were run on NuPage prefabricated Bis-Tris gradient (4-12%) gel in MES buffer (with 500 μL Antioxidant in inner chamber (~200 mL)). 200 V.
Gel loading hTERT D9 + HEK293 & HeLa -S see codes 21June2010
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Gel
cAMP prec.
| X
| X
| Invitrogen Marker
(8 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
| X
| X
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- Proteins were transferred using the NuPage system (transfer buffer +10% methanol), 30 V for 1.5 h
- All of the markers was transferred to blot, to check gel was Coomassie stained after blotting
- cAMP precipitation Ponceau S staining
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cAMP precipitation gel
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