RE: Enzyme Histochemistry for Glycogen Storage
Amy,
I'm
not up with the current train of thought on this, however the method developed
by Takeuchi (J Histochem Cytochem 1958;6:208) is said to distinguish between
phosphorylase (EC 2.4.1.1) and alpha-D glucan branching enzyme (EC 2.4.1.18).
The differentiation takes place by post-incubation treatment with alpha-amylase
and beta-amylase (branching enzyme being completekly digested with alpha-amylase
and not with beta). Also branching enzyme is inhibited by ethanol (20%) and
HgCl. I tried it out many moons ago with some success, but there was never any
demand for it so it got dropped.
The
most recent version of Pearse's "Histochemistry Vol 3" will probably have more
info, but I don't have that to hand.
Ronnie
Houston
Regional Histology Operations Manager
Bon
Secours HealthPartners Laboratories
5801
Bremo Road
Richmond, VA 23832
(804)
287 7972
Does anyone out there in histo land know of a procedure for staining
"brancher" and "debrancher" enzymes....or are these strictly metabolic tests
that have to be performed on ground up flash frozen tissue?
Amy
S.Porter, HT(ASCP)
Michigan State University
College of Human
Medicine
portera203@yahoo.com
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