To describe the formulation of both reducing and non-reducing 5X SDS-PAGE sample buffer.
Polyacrylamide gel electrophoresis in SDS containing buffers using multiphasic buffer systems: Properties of the stack, valid Rf measurement and optimized procedure. Anal. Biochem. (1978), 78:459
- Make 1M Tris pH 6.8 by adding 12.1 g Tris to 100 ml DI water. Stir to dissolve.
- Standardize pH meter to pH 4 and 7.
- pH solution to 6.8 using 37% HCl. If pH is overshot, raise pH with a 1:10 dilution of 50% NaOH/DI.
For 5X REDUCING buffer: Add 5.2 ml 1M Tris pH 6.8 to a 1 gram vial of DTT. Swirl to dissolve.
For 5X NON-REDUCING buffer: Add 5.8 ml 1M Tris pH 6.8 to 50 ml disposable centrifuge tube.
- Add 1.3 g SDS. Rock to dissolve.
- Add 6.5 ml Glycerol and 130 µl 10% Bromophenyl Blue. Rock to mix at least 30 minutes.
- Aliquot to screw cap tubes ( 24 x 0.5 ml) and store at -20°C.