Don Rio
Revised by Smadar Admon

PROTEIN GELS - SDS POLYACRYLAMIDE

Stock Solutions: (1) 30% Acrylamide;/0.8% Bis
(2) 1M Tris, pH 6.8
(3) 1M Tris, pH8.8
(4) 10% SDS
(5) 10% APS
(6) Laemmli buffer (sample buffer) (prepare fresh):
For 1 ml: 100 µl glycerol
200µl 10% SDS
125µl 1M Tris, pH 6.8
50µl BME
0.05% BPB
525µl ddH20
(7) Running buffer (for 4 liters): 57.6gr Glycine
12.1gr Tris base
4.0gr SDS
SDS polyacrylamide; protein gels;
Recipes for gels:

% running gel 5% 7% 9% 10% 12% 14% 16%
30%/0.8% AA 2.66ml 3.7ml 4.8ml 5.33ml 6.4ml 7.46ml 8.53ml
ddH20 7.1ml 6.0ml 5.0ml 4.43ml 3.36ml 2.3ml 1.23ml

Add: 6ml 1M Tris, pH 8.8
160µl 10% SDS
80µl 10% APS
8-15µl Temed

Stacking gel (5%, 5ml): 0.625ml 1M Tris, pH 6.8
0.83ml 30%/0.8% AA
3.46ml ddH2O
50µl 10% SDS
25µl 10% APS
6µl Temed

Staining Gels:

Staining Solution: 10% acetic acid
50% methanol
0.25% Coomasie brilliant blue R-250

Destaining Solution: 10% acetic acid
20% methanol