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 Project name
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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
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'
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S0+DMSO
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S0+DMSO
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S0+DMSO
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st-Ht31 (50 µM)
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st-Ht31 (50 µM)
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st-Ht31 (50 µM)
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2 |
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4 |
5 |
6
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| CTR |
A |
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| CSE (15%) |
B |
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| CSE+db-cAMP (500 µM) |
C |
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| CSE+ mPKI (400 nM) |
D |
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| 02.100810
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Control
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8Pcpt
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6Bnz
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Ht31
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Ht31+6Bnz
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DMSO
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4 |
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6
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| CTR |
A |
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| CTR |
B |
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| CSE |
C |
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| CSE |
D |
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4-12% Gradient NuPage Gel loading hTERT D9 + HEK293, HeLa -S & Jurkat see codes 21June2010
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1
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2
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3
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4
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5
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6
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7
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8
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9
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10
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11
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12
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Gel
cAMP prec.
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Invitrogen Marker
(8 μL)
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Jurkat (LYSATE)
(20 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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Results
- Ponceau staining cAMP precipitation blot (4-12% NuPage)
- Coomassie staining of the gel AFTER blot
Discussion
- AKAP9 is also not amplified in the Jurkat cells, even though these SHOULD express them. Waiting for the AKAP450 immunoblot if positive, talk to Dharmacon (coming tomorrow anyway).
Attachment
<html>
<a href="http://openwetware.org/wiki/Image:10082010_qPCR_AKAP12,_AKAP9.xls">
<img src="http://openwetware.org/images/f/f8/Owwnotebook_icon.png" alt="" width="50" height="50">
</a>
</html> qPCR Jurkat expression AKAP12, AKAP9, GAPDH
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