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RECIPES > TRICINE GEL RECIPE

> Preparation of Stock Solutions Required for Making Tricine SDS Gels

   

> Preparation of Tricine SDS Gels

   

> Preparation of the Stock Solutions Required for Running Tricine SDS Gels

    > Preparing to Run Tricine SDS Gels

Preparation of Stock Solutions Required for Making Tricine SDS Gels

  • Prepare acrylamide stock (in fume hood, wear gloves):
    • Weigh out acylamide and bisacrylamide into graduated cylinder.
    • Add H2O through 0.2 µm filter to appropriate volume.
    • The acrylamide stock is 43.65% w/v acrylamide and 1.35% w/v bisacrylamide (T = 45%
  Acrylamide stock - 250 mL Acrylamide stock - 500 mL Acrylamide stock - 1 kg acrylamide
Acrylamide 109.12 g 218.24 g 1 kg
Bisacrylamide 3.37 g 6.74 g 30.88 g
Filtered H2O

to 250 mls,

to 500 mL to 2.3 L
  • Prepare gel buffer (in fumehood):
    • In a beaker add Tris and SDS (powder) and most of the water. Then add enough HCl to get a pH of 8.45. Test with pH meter in fumehood. Finally add enough water to get the required volume.
  Gel buffer - 500 mL Gel buffer - 3 L
Tris 181.5 g 1090 g
H2O to about 400 mL  
HCl pH to 8.45 with HCl (35-40mL) pH to 8.45
20% SDS 7.5 mL 45 mL
H2O to 500 mL to 3 L

 

Reference:

Hermann Schägger and Gebhard von Jagow, " Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa", Analyt. Biochem. 166, 368-379 (1987)