Cell Lysate Preparation:Cell lysate was prepared by homogenization in modified RIPA buffer (150mM sodium chloride, 50mM Tris-HCl, pH 7.4, 1mM ethylenediaminetetraacetic acid, 1mM phenylmethyl-sulfonyl flouride, 1% Triton X-100, 0.2% sodium deoxycholate, 0.2% sodium dodecylsulfate (SDS), 5ug/ml of aprotinin, 5ug/ml of leupeptin. Cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The lysate was boiled for 5 min in 1 x SDS sample buffer (50mM Tris-HCl pH 6.8, 12.5% glycerol, 1% SDS, 0.01% bromophenol blue), 5% b-mercaptoethanol.Growth media: DMEM/Hams F12 & 10% NCS (Newborn Calf serum)Application:Suitable for use in Western Blot. Other applications not tested.Recommended Dilution:Ready to load on SDS-PAGE for Western blotting, 20ug per lane is recommended for mini gel.Storage and Stability:May be stored at -20°C for 3 months. For long-term storage, store at -70°C. Aliquots are stable for 12 months at -70°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.