Optimal detection of cholinesterase activity in biological samples: Modifications to the standard Ellman's assay

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00179906%3A_____%2F14%3A10281198" target="_blank" >RIV/00179906:_____/14:10281198 - isvavai.cz</a>

Result on the web

<a href="http://dx.doi.org/10.1016/j.ab.2014.05.031" target="_blank" >http://dx.doi.org/10.1016/j.ab.2014.05.031</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.ab.2014.05.031" target="_blank" >10.1016/j.ab.2014.05.031</a>

Result language

angličtina

Original language name

Optimal detection of cholinesterase activity in biological samples: Modifications to the standard Ellman's assay

Original language description

Ellman's assay is the most commonly used method to measure cholinesterase activity. It is cheap, fast, and reliable, but it has limitations when used for biological samples. The problems arise from 5,5-dithiobis(2-nitrobenzoic acid) (DTNB), which is unstable, interacts with free sulfhydryl groups in the sample, and may affect cholinesterase activity. We report that DTNB is more stable in 0.09 M Hepes with 0.05 M sodium phosphate buffer than in 0.1 M sodium phosphate buffer, thereby notably reducing background. Using enzyme-linked immunosorbent assay (ELISA) to enrich tissue homogenates for cholinesterase while depleting the sample of sulfhydryl groups eliminates unwanted interactions with DTNB, making it possible to measure low cholinesterase activityin biological samples. To eliminate possible interference of DTNB with enzyme hydrolysis, we introduce a modification of the standard Ellman's assay. First, thioesters are hydrolyzed by cholinesterase to produce thiocholine in the absence

Czech name

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Czech description

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Type

J_x - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)

CEP classification

FR - Pharmacology and apothecary chemistry

OECD FORD branch

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Project

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Continuities

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2014

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Analytical Biochemistry

ISSN

0003-2697

e-ISSN

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Volume of the periodical

462

Issue of the periodical within the volume

October

Country of publishing house

US - UNITED STATES

Number of pages

9

Pages from-to

67-75

UT code for WoS article

000341803700012

EID of the result in the Scopus database

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