SDS-PAGE Gel Recipes

Find SDS-PAGE recipes for stacking gel, separating gel and buffer recipes. Essential for western blotting.

In order to target proteins with MWs between 20 and 200 kDa, you will need to create a conventional SDS-PAGE gel using the recipes shown below. The percentage of gel you require corresponds with the MW of your target protein.

Recipe 1

Separating Gel (mls, total 10ml)
MW of target protein (kDa)	80-200	35-100	25-60	20-40
Gel Percentage	8%	10%	12%	15%
ddH2O	2.1	1.5	0.8	0
30% Acrylamide	2.7	3.3	4	5
2x Separating Buffer	5.0	5.0	5.0	5.0
10% APS	0.1	0.1	0.1	0.1
TEMED	0.01	0.01	0.01	0.01

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Stacking Gel (mls) 4ml
	4ml	6ml	8ml
MW of target protein (kDa)	-	-	-
Gel Percentage	4%	4%	4%
ddH2O	1.4	2.1	2.7
30% Acrylamide	0.5	0.8	1.1
2x Stacking Buffer	2.0	3.0	4.0
10% APS	0.04	0.06	0.08
TEMED	0.004	0.006	0.008

Recipe 2

2x Separating Buffer Recipe (makes 1000ml)
Tris HCl (pH 8.8)	90.8g
SDS	2.0g
Dissolve compounds thoroughly. Adjust pH slowly to pH 8.8 with concentrated HCl, then add ddH2O to 1000ml.

Recipe 3

2x Stacking Buffer Recipe (makes 1000ml)
Tris HCl (pH 6.8)	30.35g
SDS	2.0g
Dissolve compounds thoroughly. Adjust pH slowly to pH 6.8 with concentrated HCl, then add ddH2O to 1000ml.

Recipe 4

1x Running Buffer Recipe (makes 1000ml)
Tris-base	1.51g
Glycine	7.5g
SDS	0.5g
Dissolve compounds thoroughly, then add ddH2O to 1000ml.