Description

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.