Re: MS:Best stain for sub-pmole digests

Christian Lombardo (crl@burnham-inst.org)
Fri, 22 Oct 1999 08:05:44 -0700

>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

Dear Lisa,

We use a the EMBL silver staining protocol which is a modification of one
of the many silver staining methods out there. Two references where you
can find the protocol are:

Analytical Chemistry (1996) vol. 68, p. 850.
Methods in Molecular Biology volume 112, p 513. (contains an updated
version of the same protocol. This is what we use)

This protocol is very compatible with mass spectrometry analysis, both
MALDI and ESI/nanospray. For example, the gluteraldehyde cross-linking
step used in some silver staining protocols is omitted and the gel fixing
steps are much shorter. We use this protocol routinely for MALDI analysis
of tryptic in-gel digests with great results. If you'd like, I can email
you a copy of it.

-Chris-

***********************************
Christian R. Lombardo, PhD
Director, Molecular Analysis Facility
The Burnham Institute
10901 North Torrey Pines Road
La Jolla, CA 92037
858-646-3108
crl@burnham-inst.org
***********************************