SW480 cell lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylene diamine tetra acetic acid, 1 mM phenyl methyl sulfonyl flouride, 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. The cell 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% b-mercaptoethanol.
Our Abpromise guarantee covers the use of ab3957 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration. SW480 cell lysate is ready to load on SDS-PAGE for Western blotting, 10 - 20 µg per lane is recommended for mini gel.|
ab3957 has not yet been referenced specifically in any publications.