5x Western blot loading bufferΒΆ

Contributed by Martin Fitzpatrick, University of Birmingham, United Kingdom

15ml stock solution of western blot loading buffer. Dilute for use.

  • To prepare base solvent add 3ml 20% SDS to add 3.75mL 1M Tris buffer at pH 6.8 in a suitable container.
  • Add 9 mg bromphenol blue, 1.16 gm DTT (or 2.4ml B-mercaptoethanol) and mix well.
  • Add 4.5mL glycerol to the solution, mix well.
  • Make up to a final volume of 15ml with dH20 and mix again thoroughly
  • Store at 4’C. Dilute to use.