SDS-PAGE Sample Buffer for Reducing Conditions (Concentrated)

Dissolve 3.78 g of tris(hydroxymethyl)aminomethane, 10.0 g of sodium dodecyl sulfate and 100 mg of bromophenol blue in water. Add 50.0mL of glycerol and dilute to 200 mL with water. Add 25.0 mL of 2-mercaptoethanol. Adjust to pH 6.8 with hydrochloric acid, and dilute to 250.0 mL with water.

Alternatively, dithiothreitol may be used as reducing agent instead of 2-mercaptoethanol. In this case prepare the sample buffer as follows: dissolve 3.78 g of tris(hydroxymethyl)aminomethane, 10.0 g of sodium dodecyl sulfate and 100 mg of bromophenol blue in water. Add 50.0 mL of glycerol and dilute to 200 mL with water. Adjust to pH 6.8 with hydrochloric acid, and dilute to 250.0 mL with water. Immediately before use, add dithiothreitol to a final concentration of 100 mM.