galactose/galactose oxidase assay information needed

larson eric elarson at ux1.cso.uiuc.edu
Tue May 2 23:14:10 EST 1995


bipin dalmia <dalmiabk at phibred.com> writes:

>hi: i need to set-up an assay for galactose using the galactose oxidase,
>peroxidase method. would need any references to such an assay. also, does
>anyone know if a commercial kit is available with preweighed enzymes etc.
>similar to the one from Sigma for glucose assay? Sigma does not have a
>galactose kit. if you can send any information, i would appreciate very
>much. either post, or e-mail to dalmiabk at phibred.com. thanx.
=============
 
The Worthington enzyme catalog lists a procedure for assaying Galactose
Oxidase.  The book lists a review (Malmstron et al. (1975) The Enzymes,
XIIB, 3rd. Ed. (Boyer, P. D., ed.) pg 527).  
 
I suggest starting here (either the Worthington catalog or The Enzymes).
 
The procedure is fairly easy, but I don't know if anyone still produces the
kits (the Worthington catalog states they do, but it is an old catalog -- if
you've ever seen a Wothington catalog you'd understand why people save
these as reference works).
 
I've done the assay -- it's relatively straightforward (typical assay with
lots of additions).
 
I do suggest caution if you're going to have glycerol around.  There is a
very low activity with glcyerol (makes glyceraldehyde) that can futz up
long-term incubations (easy to assay this activity -- just have 20%
glycerol in your assay and you should get substantial activity without
galactose being present.

Ship me a fax number and I'll send you the two pages of the Worthington
catalog (and an ancient page of typed notes -- which I have no intention
of typing in this evening).
 
-- 
Eric Larson                  | University of Illinois at Urbana-Champaign
USDA/Agronomy                | 190 PABL; 1201 W. Gregory; Urbana, IL 61801
elarson at ux1.cso.uiuc.edu     | Voice 217.244.3079  Fax 217.244.4419
Fidonet: 1:233/4.1           | My opinions are my own, but correct :-) 



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