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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.
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