I don't have 1.5M tris HCl buffer for the resolving gel but I have 1 M tris HCl buffer with pH 8.8 will that work for SDS PAGE? do I need to change the amount added to the mix?
If the recipe includes water, you can reduce the volume of water and increase the volume of Tris buffer to get to the same final concentration.
thank you for your response, please elaborate Adam B Shapiro
Here is an example of a 10% resolving gel recipe:
H2O 4.1 ml
Acrylamide/bis (30% 37.5:1) 3.3 mL
Tris–HCl (1.5 M, pH 8.8). 2.5 mL
SDS 10%. 100 µL
N,N,N′,N′-tetramethylethylene-diamine (TEMED) 10 µL
Ammonium persulfate (APS), 10% 32 µL
The total volume is 10 mL, of which 2.5 mL is 1.5 M Tris, so the final Tris concentration is (2.5/10) x 1.5 M = 0.375 M
If you only have 1 M Tris, to make 10 mL of 0.375 M, you need 10 x (0.375/1) = 3.75 mL.
So, instead of using 2.5 mL of 1.5 M Tris and 4.1 mL of water, you can use 3.75 mL of 1 M Tris and 2.85 mL of water.
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