Glucose Oxidase

From:Chris van der Loos

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Luis Chiriboga wrote:

Date: 17 Oct 2002 11:07:15 -0500
From: Luis Chiriboga 
Subject: Glucose Oxidase

Hi all
Has anyone ever tried to detect Glucose oxidase enzyme activity directly in
FF-PE tissues???
Any suggestions would be helpful


Dear all,
The application of GOX from Aspergillus niger as marker enzyme is=20
theoretically ideal since the GOX enzyme is not present in mammals. During 
the 70-80ties GOX as enzymatic marker was even commercially available! 
However, in my hands and that of many others background of unknown origin 
occurred. As far as I am aware of nobody is using GOX anymore.
For staining protocol see: Campbell and Bhatnagar, J Histochem Cytochem 
24:448 (1976) and Clark et al J Histochem Cytochem 30:27 (1982)

After removing some dust the following protocol appeared:
Tris-HCl buffer (0.1 M, pH8.3)			5.0 ml
b-D(+) glucose (Sigma G5250)			67 mg
NBT (Sigma N6876) 5 mg/ml			1.34 ml
pPMS (Sigma P9625) 1mg/2 ml dist. water	0.34 ml (keep in dark!)
dist. water					3.33 ml

Incubate at 37C in the dark for 30-60 min
wash with distilled water
mount aqueously	

Success,
Chris
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