Document Detail


Development of a microplate fluorescence assay for kynurenine aminotransferase.
MedLine Citation:
PMID:  21059337     Owner:  NLM     Status:  In-Process    
Abstract/OtherAbstract:
Inhibition of kynurenine aminotransferases (KATs) is a strategy to therapeutically reduce levels of kynurenic acid (KYNA), an endogenous antagonist of glutamatergic N-methyl-D-aspartate (NMDA) and cholinergic α₇ nicotinic receptors. Several methods of measuring KAT activity in vitro have been developed, but none is well-suited to high throughput and automation. In this article, we describe a modification of existing high-performance liquid chromatography (HPLC)-based methods that enables the development of a 96-well microplate assay in both enzyme- and cell-based formats using human KAT I as an example. KYNA enzymatically produced from L-kynurenine is measured directly in a reaction mixture fluorimetrically.
Authors:
Jacky Wong; William J Ray; Anna Y Kornilova
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Publication Detail:
Type:  Journal Article     Date:  2010-11-06
Journal Detail:
Title:  Analytical biochemistry     Volume:  409     ISSN:  1096-0309     ISO Abbreviation:  Anal. Biochem.     Publication Date:  2011 Feb 
Date Detail:
Created Date:  2010-12-23     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  183-8     Citation Subset:  IM    
Copyright Information:
Copyright © 2010 Elsevier Inc. All rights reserved.
Affiliation:
Neurology Department, Merck, West Point, PA 19486, USA.
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