Re: Sulfur staining??
There are several ways to stain -SH groups, of
varying sensitivity and specificity. Two that I
have used and would recommend are:
1. Acidified permanganate followed by alcian
blue, pH 1 or lower. Rather low sensitivity,
so there's little background, but sulphated
mucosubstances also stain (do a control
alcian blue without prior oxidation).
This the method of Adams & Sloper 1955
Lancet I:651-652, but with permanganate
oxidation instead of performic acid, which
is nasty stuff. I think the first publication
of the modification was Pasteels & Herlant
1962 Z. Zellforsch. 20-39.
2. The ferric ferricyanide reaction. High
sensitivity, but other reducing groups
also react (see Lillie & Donaldson 1974
Histochem. J. 6:679-684). The prussian blue
product fades with time (a few years) in
some mounting media, so take photos.
Another simple reagent is mercurochrome
(20 mg in 0.5ml water, then add 100 ml
of N,N-dimethylformamide). Stain 1 hr
for green fluorescence, or 48 hr for red
staining of -SH groups. After staining
wash in 3 changes of DMF, 2 each of ethanol
& xylene, and mount with a non-fluorescent
Lillie & Fullmer's 1976 textbook is a good
source for methods of this kind.
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London, Canada N6A 5C1
> I was wondering if anyone knew of a stain that would target SH groups.
> I'm trying to stain hyaluronan that's been thiolated and I'm finding
> that alsian blue is very faint. Is there a way to go after the sulfur
> Owen Lewis
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