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

 .todo { color: red } .done { color: green} {| width="800"
 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] ELISA, qPCR Jurkat, SDS-PAGE cAMP precipitation
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


 * colspan="2"|
 * colspan="2"|

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
 * SDS-PAGE
 * qPCR Jurkat RNA

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
   [[Media:10082010_qPCR_AKAP12,_AKAP9.xls|qPCR Jurkat expression AKAP12, AKAP9, GAPDH]]


 * }