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Akrylamide Gels


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#1 Sly

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Posted 24 August 2010 - 01:47 AM

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.

#2 tfitzwater

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Posted 24 August 2010 - 07:01 AM

Bisacrylamide is not soluble much above 2%

#3 mdfenko

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Posted 24 August 2010 - 09:55 PM

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.
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