Full Name: Sea star (Pisaster ochraceus) oocyte - 1-methyladenine-treated - Whole lysate

Product Type Specific: Whole lysate

Product Description: Whole GVBD-positive, maturing sea star oocyte lysate

Product Method: Oocytes teased from ripe seastar oocyte gonads were resolved from follicle cells by 3 rounds of centrifugation at 2500 rpm for 5 min each time. Lysates were prepared from isolated oocytes 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.

Storage Conditions: -70°C

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