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

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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)