4x SDS Protein Sample Buffer

SDS protein sample buffer is added to solubilized protein samples prior to analysis by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). 1 part of this 4x buffer is added to 3 parts solubilized protein. Alternatively, the buffer can be diluted to 1x (with water) and added directly to pellets of bacterial or eukaryotic cells. In either case, the mixture should be heated at 95-100 C for 5 minutes to denature proteins prior to SDS-PAGE.

Final concentrations:

  • 40% Glycerol
  • 240 mM Tris/HCl pH 6.8
  • 8% SDS
  • 0.04% bromophenol blue
  • 5% beta-mercaptoethanol

For 10 ml:

  • 4 ml 100% glycerol
  • 2.4 ml 1M Tris/HCl pH 6.8
  • 0.8 g SDS
  • 4 mg bromophenol blue
  • 0.5 ml beta-mercaptoethanol
  • 3.1 ml H2O

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