A High-Throughput Colorimetric Microplate Assay for Determination of Plasma Arginase Activity.

Autor:	Smith NJ; University of Sydney, Camperdown, NSW, Australia., Maddahfar M; University of Technology Sydney, Ultimo, NSW, Australia., Gunasegaran B; University of Sydney, Camperdown, NSW, Australia., McGuire HM; University of Sydney, Camperdown, NSW, Australia. Helen.mcguire@sydney.edu.au., Fazekas de St Groth B; University of Sydney, Camperdown, NSW, Australia.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2023; Vol. 2620, pp. 273-286.
DOI:	10.1007/978-1-0716-2942-0_29
Abstrakt:	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. (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze:	MEDLINE
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