User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/04/27

Summary

• RII overlay
  • Activate membrane in EtOH 100% for 15 sec.
  • Rinse with H₂O
  • Block in Blocking buffer for 3 hours
    • Blocking buffer (5% Skim milk, 0.1% BSA, 0.0001% Sodium Azide in PBS)

• RIPA BUFFER (100 mL)
  • 50 mM Tris-HCl (5 mL 1 M Tris pH 7.4)
  • Adjust to pH 7.4
  • 150 mM NaCl (0.87 g NaCl)
  • 1 mM EDTA (37.22 mg EDTA)
  • 1% Triton x-100 (1 mL Tritox x-100)
  • 1% Sodium deoxycholate
  • 0.1% SDS (1 g SDS)