Hi,
Staining SDS-PAGE gel should be decided by what you want to do after
staining a gel. If you wish to identify proteins with mass spectrometry,
MALDI or ESI, you should consider staining methods that are compatible with
those techniques.
Staining Dyes
1. Coomassie Brilliant Blue R or G or Colloidal Gold --- detection limit,
1.0 ug/spot or band, compatible with mass spectrometry
2. Imidazole-zinc --- 10 ng/spot or band, reverse phase staining: proteins
will be white and background will be dark gray, compatible with mass
spectrometry. a home made procedure is available.
3. Silver --- 1-10 ng/spot or band, compatible with mass spectrometry, a
home made procedure is available or quite numbers of commercial products are
available. I like NOVEX Silver Xpress (Now it is merged to Invitrogen).
4. Fluorescents:
SYPRO Red/Orange --- 10 ng/spot or band, compatible with mass spectrometry.
A good description for Red is published. (J. Mass Spectrometry, V35:672-682,
2000. Israel Valdes, et al)
Nile Red --- similar detection limit to Coomassie but compatible with
N-terminal sequencing or Western Blotting. It is a non-covalent and speedy
staining dye.
5. Isotope labeling
For the most sensitive and dependable stain of 50 ng protein, I vote Silver
for analyzing proteome and Sypro Red for mass spectrometry. You have to
deal with UV for fluorescent dyes, though.
KeeHong Kim
Chemical Engineering
Colorado State University
Ft. Collins, CO80521-1370
khkim@lamar.colostate.edu
970-491-1192
----- Original Message -----
From: "Norman Watts" <WATTS@calvin.niams.nih.gov>
To: "Recipients of ABRF List" <abrf@aecom.yu.edu>
Sent: Wednesday, December 13, 2000 6:14 AM
Subject: Misc, SPS-PAGE, sensitive protein stain needed
> Hi all,
>
> What is currently considered to be the most sensitive and
> dependable stain for detecting protein bands on SDS-PAGE
> gels? I expect ca. 0.05 ug/band.
>
> norman
>
> -------------------------------------------------------
>
> Norman Watts, PhD.
> Lab. of Structural Biology
> Building 6, Room B2-34
> 6 Center Drive MSC2717
> NIAMS, National Institutes of Health
> Bethesda, MD 20892-2717
> U.S.A.
> E-mail: watts@calvin.niams.nih.gov
> Phone: 301-402-3418
> Fax: 301-480-7629
>
> -------------------------------------------------------
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