A549 Whole Cell Lysate (Human Lung Carcinoma)
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DetailsCell Line A549 (Human Lung Carcinoma) Growth Media DMEM and 10% NCS (Newborn Calf Serum) Preparation A549 lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl fluoride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecyl sulfate, 5 μg/ml of aprotinin, 5 μg/ml of leupeptin. Cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. Host Human Concentration ≥ 2.0 mg/ml Formulation The lysate was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecyl sulfate, 0.01% bromophenol blue) containing 5% ß-mercaptoethanol. Expiration Date One year from date of receipt Storage This lysate should be aliquoted in working amounts and may be stored for up to one month at -20°C <b>in a manual defrost freezer. Avoid Repeated Freeze Thaw Cycles.</b> For longer term storage, freeze working aliquots at -70°C. Applications and Suggested Working Dilutions This lysate is for use in Western blotting, 10 µg to 20 µg per lane is recommended for mini gel. |
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