Re: MS:Best stain for sub-pmole digests

L. Ericsson (ericsson@u.washington.edu)
Fri, 22 Oct 1999 06:59:41 -0700 (PDT)

Hi Lisa and all,

Have you considered not staining at all when you know you have protein on
your gel? Just slice your gel into small pieces, pixels?, keeping a
record of the location and do in-gel digests on all those "pixels"
followed by LC-MS or MALDI. This was a suggestion from David Arnott from
Genentech at a recent Asilomar conference. Ag does not stain all proteins
equally and at very low levels not at all.. Not staining can also reduce
the contamination level.

-Lowell H. Ericsson, Dept. of Biochemistry, U. of Washington, Seattle, WA

On Thu, 21 Oct 1999, lisa bibbs wrote:

> What is the best stain for sub-pmole amounts of protein to be in-gel
> digested followed by Mass spec identification (MALDI followed by
> LC-MS)? I have heard that Silver staining can be very bad and
> produce no MALDI results. We haven't gone to sub-pmole yet so we
> have just been using Coomassie and such.
>
> Thanks for your help.
>
> Lisa
> Lisa Bibbs, Ph.D.
> 10550 N. Torrey Pines Rd.
> SP3
> La Jolla, CA 92039
>
> http://corefac.scripps.edu
>
>