COMT reaction using erythrocytes

Ilonka Guenther guenther at cvax.psi.ch
Tue Sep 6 01:27:48 EST 1994


Dear Netters,

I would like to establish an assay for erythrocyte Catechol-O-methyltransferase
(COMT) activity in human and primate blood. I tried it by following a
protocol from Bates et al. (Clin. Chim. Acta 94, p63-71, 1979) using EGTA and
SAM in the proposed concentrations. As a substrate I want to use [18-F]-FDOPA,
controlling the reaction by HPLC analysis and an on-line gamma detector.
I incubate the reaction mixture for 30 min at 37 degrees Celsius. 
So far I have never seen any methylation product, even though I know the 
retention time of the product.

Is there any suggestion why the reaction doesn't work?

Is there a newer protocol existent?

Any help would be appreciate very much. 

Please respond to my e-mail address.

Thanking in advance.

		Ilonka Guenther
		Guenther at cvax.psi.ch




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