2 replies to this topic

[Sly]

Hello everyone.

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.

Best regards.

[tfitzwater]

Bisacrylamide is not soluble much above 2%

[mdfenko]

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.