SDS-PAGE gel solutions

Gel Buffer

1.5M Tris, pH 8.8 with 0.4% SDS

92.5g Tris base

400 mL ddH20

pH to 8.8 with concentrated HCl (about 11mL)

add ddH2O to 500mL and recheck pH

add 2g SDS

filter and store at 4 for 6 months

Stack Buffer

0.5M Tris, pH 6.8 with 0.4% SDS

6g Tris

75mL ddH2O

pH to 6.8 with concentrated HCl (about 4 mL, then go slow with 1mL 1M)

add ddH2O to 100mL and recheck pH

add 0.4g SDS

filter and store at 4 for 6 months

SDS-PAGE 2X sample buffer

2.5mL stacking buffer

2mL 10% SDS

2.5mL 80% glycerol

2mL ddH2O

1mL Beta-Mecaptoethanol

Add a pinch of bromphenol blue to color

Aliquot and store at –20 for 6 months

Running buffer:

6g Tris base

28.8g glycine

1g SDS

per liter ddH2O

Gel Stain:

0.1% Coomassie (r-250) in 50% methanol, 10% acetic acid

Destain:

5% methanol, 10% acetic acid