Full Name: Human epidermoid carcinoma HeLa cells - PMA-treated Whole lysate

Alias: Human epidermoid HeLa cells - TPA-treated Whole lysate

Product Type Specific: Whole lysate

Product Description: Whole lysate from HeLa cells that were cultivated to 70-90% confluency and deprived of serum for 18-20 hours and then 100 nM phorbol 12,13-myristate acetate (PMA) for 5 minutes prior to harvesting

Product Method: HeLa cell lysates were cultivated to 70-90% confluency and deprived of serum for 18-20 hours and then 100 nM phorbol 12,13-myristate acetate (PMA) 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 A. 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. 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.

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