western blotting操作手册

Running Protein Gels

Solutions

10X Running Buffer (0.25 M Tris, 1.92 M glycine, 1% SDS)

121 g Tris

577 g glycine

40 g SDS

ddh20 to 4 L (check pH at 1:10 dilution (pH=~8.8)).

5X Sample Buffer (0.3125 M Tris pH 6.8, 10% SDS, 50% glycerol, 0.005% bromophenol blue, 25% 2-mercaptoethanol)

1.56 ml 2M Tris pH 6.8

1 g SDS

5 ml glycerol

0.2 ml 0.25% solution of bromophenol blue

2.5 ml 2-mercaptoethanol

ddH20 to 10ml.

30:0.8 Acrylamide:bis

0.5 M Tris pH 6.8

10% SDS

10% APS

TEMED

3 M Tris pH 8.8