Full Name: Human epidermoid carcinoma A431 cells - Whole lysate

Product Type Specific: Whole lysate in SDS-PAGE sample buffer

Product Description: Whole cell lysate boiled in SDS-PAGE sample buffer prepared from A431 cells that were cultivated to 70-90% confluency and deprived of serum for 18-20 hours and then treated for 100 ng/ml epidermal growth factor (EGF) for 5 minutes prior to harvesting

Product Method: A431 cells were cultivated to 70-90% confluency and deprived of serum for 18-20 hours and then treated for 100 ng/ml epidermal growth factor (EGF) for 5 minutes prior to harvesting. Lysates were prepared from scrapped cells that were homogenized by sonication in buffer formulated with 1% Triton X-100, 60 mM ß-glycerophosphate, pH 7.2, 20 mM MOPS, 20 mM sodium pyrophosphate, 30 mM sodium fluoride, 5 mM EDTA, 3 mM benzamidine, 2 mM EGTA, 1 mM sodium orthovanadate, 1 mM phenylmethylsulfonylfluoride, 1 mM dithiothreitol, 10 µM leupeptin, and 5 µM pepstatin. Lysates were prepared following sonication and 30 min ultracentrifugation at 100,000 rpm. Lysates were further diluted in SDS-PAGE sample buffer at a final concentration of 2 mg/ml.

Storage Conditions: -70°C

Lysate Use Description: For testing antibodies by immunoblotting.