User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/07/06

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 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] cAMP precipitation gel
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


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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.
 * 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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