|
WESTERN BLOT PROTOCOL SOLUTIONS
1) 30% Acrylamide, 37.5:1 Diluted per BMB.
2) 4X LGB 1.5 M TRIS pH 8.8, 0.4% SDS
per 500 ml 90.75 grams TRIS 10 ml 20% SDS pH to 8.8 with HCL. Dilute to 500 ml.
3) 4X UGB 0.5 M TRIS pH 6.8, 0.4% SDS
per 500 ml 60.5 grams TRIS 10 ml 20% SDS pH to 6.8 with HCL. Dilute to 500 ml.
4) 69% Glycerol. 50 ml stock.
5) 10% Ammonium persulfate. 0.1 grams in 1 ml ddH20. (make fresh q week)
6) 1OX Running Buffer
1L
0.25 M TRIS 30.26 grams
1.97 M Glycine 144.2 grams
1.0 % SDS 50 ml of 20% SDS stock
pH will be 8.3 +/- 0.2 after dilute to 1X. Don't measure pH
of 1OX stock. -
7) SDS SAMPLE BUFFER 2X: 0.125 M TRIS pH 6.8, 4% SDS, 20 %
Glycerol, 10% 2-ME, 0.02% BPB
4X UGB 2.5
20% SDS 2.0
100% Glycerol 2.0 Dilute some 1:1 with ddH20 to make
2-mercaptoethanol 1.0 1X Buffer, to run in newly poured
1% bromphenolblue 0.2 gel to define wells.
ddH20 2.3
10.0 ml
|
