SDS-PAGE Gel Recipes

SDS-PAGE Gel Recipes

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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

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.