IUBio

[Protein-analysis] Re: protein contamination with DNA???

Amanda Gillespie gillespie.amanda at gmail.com
Fri Mar 24 14:41:12 EST 2006


Yes, coomassie is incredibly insensitive to proteins - there are much more
sensitive stains out there which will confirm if you really only have one
protein.

Silver is very sensitive, but will also stain DNA, and is quite laborious.
Sypro Ruby is an easy stain, but quite expensive
Imidazole-SDS zinc staining is my method of choice - as sensitive as silver
and Sypro, but cheap, and 15 min from start to finish.

I can send you a reference for a protocol if you need one...

Amanda

On 22 Mar 2006 09:18:07 GMT, kaj.stenberg at helsinki.fi.invalid <
kaj.stenberg at helsinki.fi.invalid> wrote:
>
> nithya <blueylife at gmail.com> wrote:
>
> > Amanda Gillespie wrote:
> >> what have you stained your SDS-PAGE gel with that tells you there is
> only
> >> one band?
> >>
> >> Amanda
> >>
>
> > I have stained it with coomassie blue and i used marker as a known
> > protein of same molecular weight.
> > Thanking you for any suggestions
>
> 1) Coomassie doesn't stain all proteins well. You could try silver
> staining
> 2) Make a symmetric gel, cut in half, stain one half with coomassie or
> silver, other half with EtBr.
>
> --
> Kaj
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>



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