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Summary
- AKAP250 immunoprecipitation
- Gradient (4-12%) SDS-PAGE (NuPage)
Gel loading hTERT D9 + HEK293 & HeLa -S see codes 21June2010
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Gel
cAMP prec.
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X
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X
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Invitrogen Marker
(8 μL)
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HeLa -S
(20 μL)
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HEK293
(20 μL)
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Biorad Marker
(8 μL)
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C1
(20 μL)
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C2
(20 μL)
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C3
(20 μL)
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S1
(20 μL)
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S2
(20 μL)
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S3
(20 μL)
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X
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X
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X
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Notes
- HeLa cells had less sample put too gel, both HEK293 and HeLa are very slimy
- Forgot to lyse the ordered HEK293 cells
- Blot was blocked ON in 1% BSA in TBS-T
- Gel was Coomassie stained ON
Results
- Overnight autoradiograph of RII overlay did not change anything of the image of yesterday
- Ponceau of cAMP pull down SDS-PAGE
Discussion
- Ponceau shows bands at higher altitude, maybe look for AKAP-Lbc next to AKAP250?
- Anoying that both markers are not running equal on every height, but HiMark at least has nice sharp band @ 160 kDa on NuPage gels. Although transfer was less perfect
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