Ronnie - Thank you very much for your reply. If you can believe it we actually have an ancient text in our laboratory that had this procedure. I could'nt believe that there wasn't something simpler out there!! Since you have actual knowledge of this protocol does this cover the branching segment and the debranching segment? I think that is what I am getting from your response, just want to double check. This is something our neuropathologist just asked us to look into so i am not really up to speed on glycogen storage diseases other than the fact there are many of them. I really appreciate your help.
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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