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SDS Gel Dyes & Buffers

Posted on January 24, 2019 by admin • 0 Comments

Recipes for running SDS protein gel electrophoresis, SDS sample loading dye, and Coomassie staining protein gels.

5x SDS loading dye

125mM Tris-HCl, pH 6.75
20% (v/v) glycerol
10% (v/v) 2-mercaptoethanol
4% SDS
0.02% bromophenol blue

5x Running buffer (1L)

72 g glycine
15 g Tris-HCl
5 g SDS

Gel Stain

2.5g/L Coomassie Blue
45% (v/v) methanol
45% (v/v) glacial acetic acid

Gel Destain

10% (v/v) acetic acid
10% (v/v) methanol

Single Percentage Polyacrylamide Gels

Posted on January 24, 2019 by admin • 0 Comments

Recipe for casting common single percentage protein polyacrylamide gels. Separation Gel (10mL) 7% 10% 12.5% 15% Stacking Gel (4mL) H2O 5.1 mL 4.1 mL 3.3 mL 2.5 mL 2.1 mL Acrylamide/bis (30%) 2.3 mL 3.3 mL 4.1 mL 5.0 mL 0.8 mL Tris-HCL (1.5M, pH 8.8) 2.5 mL 2.5 mL 2.5 mL 2.5 mL 1… Continue Reading