Total Protein Cell Lysis Buffer is a nonionic detergent lysis buffer that enables efficient solubilization of the plasma and intracellular membranes, breaks weak intermolecular bonds, and solubilizes most proteins. Total Protein Cell Lysis Buffer reduces protein denaturation, protein complex disruption and loss of enzymatic activity since it contains less detergent than a typical RIPA lysis buffer. Simply, resuspend cells in Total Protein Cell Lysis Buffer and vortex vigoursly. Incubate the cell suspension at 4 ºC with shaking for 30 minutes. Vortex vigoursly for 30 seconds and pellet cellular debris. The supernatant contains the total protein lysate.
Total protein from K562 cells probed for HSF-1 expression. Total protein was isolated from K562 cells using AMRESCO’s Total Protein Cell Lysis Buffer (M329) and then resolved on a 12.5% Fluorescent SPRINT NEXT GEL® for 30 minutes at 300 V. The protein was visualized by UV transillumination (lane 1) and then transferred to PVDF. RapidBlock™ Solution (M325) was used for blocking and as the antibody diluent with HSF-1 antibody (lane 2). The blot was developed using VisiGlo Plus™ HRP Chemiluminescent Substrate Kit (N219).
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