Full Name: Human Embryonic Kidney-293 cells - Forskolin-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 HEK293 cells that were cultivated to 90% confluency and deprived of serum for 18 hours and then treated with 150 nM forskolin for 15 minutes prior to harvesting

Product Method: HEK293 cells were cultivated to 80% confluency and deprived of serum for 18 hours and then treated with 150 nM forskolin 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.