Protocol for measuring erythrocyte glutathione reductase activity coefficient to assess riboflavin status

Damon A Parkington; Albert Koulman; Kerry S Jones

doi:10.1016/j.xpro.2023.102726

Protocol for measuring erythrocyte glutathione reductase activity coefficient to assess riboflavin status

STAR Protoc. 2023 Dec 15;4(4):102726. doi: 10.1016/j.xpro.2023.102726. Epub 2023 Nov 20.

Authors

Damon A Parkington¹, Albert Koulman², Kerry S Jones³

Affiliations

¹ Nutritional Biomarker Laboratory, MRC Epidemiology Unit, University of Cambridge, Cambridge CB2 0SL, UK. Electronic address: damon.parkington@mrc-epid.cam.ac.uk.
² Nutritional Biomarker Laboratory, MRC Epidemiology Unit, University of Cambridge, Cambridge CB2 0SL, UK.
³ Nutritional Biomarker Laboratory, MRC Epidemiology Unit, University of Cambridge, Cambridge CB2 0SL, UK. Electronic address: kerry.jones@mrc-epid.cam.ac.uk.

Abstract

Riboflavin (vitamin B2) is a component of the co-enzyme flavin adenine dinucleotide (FAD). The activity coefficient of erythrocyte glutathione reductase (EGRAC), a FAD-dependent enzyme, is a biomarker of riboflavin status. Here, we describe a protocol for measuring unstimulated (basal) and FAD-stimulated (activated) erythrocyte glutathione reductase activity to calculate EGRAC. We describe the steps for preparing washed red blood cells and hemolysates; preparing reagents; loading, incubating, and reading the 96-well plate; and calculating the results. For complete details on the use and execution of this protocol, please refer to Hess et al.¹.

Keywords: Chemistry; Health Sciences.

MeSH terms

Erythrocytes
Flavin-Adenine Dinucleotide*
Glutathione Reductase
Riboflavin*

Substances

Flavin-Adenine Dinucleotide
Glutathione Reductase
Riboflavin