Preparation: Cell lysate was prepared in modified RIPA buffer (50mMTris, pH7.2, 150mM NaCl, 1mM EDTA, 1mM EGTA, 1% NP-40 , 1mM Na3VO4, 5mM NaF, 400uM PMSF, 1ug/ml of pepstatin A, 5ug/ml of aprotinin, 5ug/ml of leupeptin). Cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The cell lysate was diluted and boiled for 5 min with nonreducing sample buffer (50 mM Tris-HCl, pH 6.8, 5% glycerol, 1% SDS, 0.002% bromphenol blue) containing 5% beta-mercaptoethanol.

Concentration: 2 mg/ml

Storage: Stable for 6 months at -20oC from date of shipment. For maximum recovery of product, centrifuge the original vial after thawing and before removing the cap. Aliquot to avoid repeated freezing and thawing.

Application: Cell lysate is ready to load on SDS-PAGE for Western blotting, 20 µg per lane is recommended for mini gel.