Full Name: Human breast carcinoma MCF7 cells - Insulin-treated - Cytosolic lysate

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

Product Description: Cytosolic lysate boiled in SDS-PAGE sample buffer prepared from MCF7 cells that were cultivated to 80% confluency and deprived of serum for 18 hours and then treated with 10 µg/ml insulin for 15 minutes prior to harvesting

Product Method: MCF7 cells were cultivated to 80% confluency and deprived of serum for 18 hours and then treated with 10 µg/ml insulin for 15 minutes prior to harvesting. Lysates were prepared from scrapped cells that were homogenized by sonication in buffer formulated with 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 A. Cytosolic 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.