HELP: peptide separation on SDS-PAGE

D.K. dk at no.email.thankstospam.net
Wed Nov 28 08:13:11 EST 2001


 "L. Lintott" <llintott at shaw.ca> wrote:
>With older BioRad mini's this can be a
>problem as they tend to leak from the upper chamber.  

Not if they are properly assembled. 

>I always put lots
>of grease on the gasket to prevent this.

I thought the whole idea of their design was to get rid
of disgusting grease :-) 

>I also use a polypetide fixing solution on the gel before I stain
>coomassie (40% methanol and 10% acetic acid).  

Does not work well for many polypeptides below 5k. Aldehyde
fixation is a lot more pain but works better. 

>Recently I have
>switched to using Invitrogen's "Simply Blue" and find that it works very
>well for the smaller proteins, and is somewhat simpler to use. 

It is simply Bradford's reagent. You can make 1 L of it in ten minutes
for the price of 60 mg Coomassie and 100 ml conc. phosphoric acid. 

DK




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