Akrylamide Gels - (Aug/24/2010 )
I have a question about akrylamide gels for western blot. I'm trying to prepare a gel, old fasioned style.
In my very old protocol that I'm using, it says 30% akrylamide and 8% Bisakrylamide. However, we have a pre-made solution, made by BIO Rad. On the bottle it says 30% akrylamide/Bis 29:1 (3.3% C).
My question is, how important is the different concentrations? Would it make a difference in my end result?
I should mention that my experiement is for screening purpose, so I'm not trying to detect any particular protein.
I'd be very thankful if someone could shed a light on the matter.
Bisacrylamide is not soluble much above 2%
the bisacrylamide is probably 0.8%. the ratio is 37.5:1 (not 29:1). the difference in bisacrylamide concentration will make a difference in the appearance of the final result.