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

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