IUBio

Suggestions: Peptide dyes for hydrophobic amino acids.

Tom McCloud McCloud at dtpax2.ncifcrf.gov
Wed Jul 15 11:40:45 EST 1998


I personally do not work in this area, but asked a guy in a different lab for
suggestions.    His first choice would be use of a dye called spyro orange
(BioRad or Molecular Probes?) which interacts with the SDS rather than the
pepetide.  It is fluorescent and you need a laser densitometer to
detect/quantify.  Very sensitive if it will work.

Second suggestion: Procion Blue (Sigma), which binds covalently to proteins
(as opposed to Coomassie which is not covalent), but we don't know which amino
acids it reacts with.  Probably not a real sensitive method.

Third suggestion: colloidal gold stain. BioRad or Amersham sell kits used to
do western blotting.   Usually done on a membrane: probably have to do some
modification to adapt to acrylamide gels.  Should be very sensitive if it
works.

Sorry I am not more familiar with the techniques, but if you need more
information I will try to put you and the protein guy in direct contact.
Tom McCloud   SAIC/Frederick Cancer Research

Doug Stemke wrote:

> A graduate student in our department who is synthesizing a hydrophobic
> peptides has run into an interesting problem.  He has found that since he
> has no charged residues on his peptides he is unable to stain his peptides
> in SDS gels with either silver stain or Coomassie blue.  He has been doing
> some negative staining on his gels which work ok, but he is unable to
> effectively quantitate his results.  Does anyone here have a
> recommendation for a stain that would effectively bind hydrophobic amino
> acids in SDS gels?
>
> Thanks.
>
> Doug
>
> ******************************************************************
> Douglas J. Stemke, Ph.D.
> Biological Process Technology Institute
> University of Minnesota
> St. Paul, MN
> dstemke at biosci.cbs.umn.edu
> ******************************************************************






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