C2C12 Cell Lysate, Murine

C2C12 cells are a useful model to study the differentiation of non-muscle cells to skeletal muscle cells (e.g myosin phosphorylation mechanisms) and express muscle proteins and the androhen receptor (AR). Murine C2C12 cell lysate was prepared by homogenization in modified RIPA buffer (50mM Tris-HCl, pH 7.4, 1% Triton X-100, 0.2% sodium deoxycholate, 0.2% sodium dodecylsulfate (SDS), 1mM sodium ethylenediaminetetraacetate, 1mM phenylmethylsulfonyl flouride, 1 ug/ml of aprotinin, 5ug/ml of leupeptin). Cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The lysate was boiled for 5 min in 1X SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% SDS, 0.01% bromophenol blue) containing 5% b-mercaptoethanol. \n\nNote: Cell line-C2C12 (muscle; myoblast); Growth media-DMEM & 10% fetal bovine serum.\n\nApplications: \nSuitable for use in Western Blot. Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: 20ug per lane, mini gel.\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nAliquot to avoid repeated freezing and thawing and freeze at -70 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Aliquots are stable for at least 12 months.

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