User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/07/08

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


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Summary

 * * RII overlay, washing of membranes and taking pictures
 * Other membranes looked okay, for my membrane see below

Results

 * RII Overlay 3h autoradiograph
 * Only RII subunits seem to be visible, HEK293 has a band around 66 kDa which should at least be >75 kDa (AKAP79)

Discussion

 * Maybe a new lysis buffer should be better?
 * RIPA BUFFER GRONINGEN
 * 40 mM Tris
 * 150 mM NaCl
 * 1% Igepal CA-630
 * 1% deoxycholic acid
 * pH 7.4
 * Addition
 * 1 mM NaF
 * 1 mM Na3VO4
 * 10 µg/ml aprotinin
 * 7 µg/ml leupeptin
 * 7 µg/ml pepstatin A
 * 1 mM PMSF


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