Full Name: Human Embryonic Kidney-293 cells - Forskolin-treated - Cytosolic lysate

Product Type Specific: Cytosolic lysate

Product Description: Cytosolic lysate 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 homogenizing buffer at a final concentration of 3 mg/ml.

Storage Conditions: -70°C

Lysate Use Description: For testing antibodies by immunoprecipitation or immunoblotting, and for assays of enzymes.