A high-throughput colorimetric microplate assay for determination of plasma arginase activity

Natalie J. Smith, Mahnaz Maddahfar, Bavani Gunasegaran, Helen M. McGuire, Barbara Fazekas de St Groth

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

Arginase, an enzyme involved in the urea cycle, is gaining attention as a critical player in numerous chronic pathologies. Additionally, increased activity of this enzyme has been shown to correlate with poor prognosis in a range of cancers. Colorimetric assays that measure the conversion of arginine to ornithine have long been used to determine the activity of arginase. However, this analysis is hindered by a lack of standardization across protocols. Here, we describe in detail a novel revision of the Chinard’s colorimetric assay used to determine arginase activity. Dilution series of patient plasma are plotted to form a logistic function, from which activity can be interpolated by comparison to an ornithine standard curve. Inclusion of patient dilution series rather than a single point increases the robustness of the assay. This high-throughput microplate assay analyzes 10 samples per plate to produce highly reproducible results.
Original languageEnglish
Title of host publicationProtein arginylation
Subtitle of host publicationmethods and protocols
EditorsAnna S. Kashina
Place of PublicationNew York
PublisherThe Humana Press
Chapter29
Pages273-286
Number of pages14
Edition2
ISBN (Electronic)9781071629420
ISBN (Print)9781071629413
DOIs
Publication statusPublished - 18 Apr 2023
Externally publishedYes

Publication series

NameMethods in Molecular Biology
PublisherSpringer
Volume2620
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Arginase
  • Arginine
  • Colorimetric assay
  • Enzyme activity
  • Microplate

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