[Protein-analysis] adding nucleases to SDS PAGE samples
da8688 at yahoo.com
Sat Mar 25 05:27:25 EST 2006
What are the general opinions of the group regarding
digesting tissue culture cell preparations with
nucleases during sample prep for SDS-PAGE. If one
doesn't add nucleases, are there physical or chemical
strategies to make samples less viscous?
Thanks for any thoughts.
--- Amanda Gillespie <gillespie.amanda at gmail.com>
> Yes, coomassie is incredibly insensitive to proteins
> - there are much more
> sensitive stains out there which will confirm if you
> really only have one
> 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
> I can send you a reference for a protocol if you
> need one...
> 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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> If at first you don't succeed, destroy all evidence
> that you tried --- Homer
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