An enzymatic colorimetric assay for glucose-6-phosphate

作者：Zhu Aiping; Romero Roberto; Petty Howard R^*

来源：Analytical Biochemistry, 2011, 419(2): 266-270.

DOI：10.1016/j.ab.2011.08.037

• WST-1
• Glucose-6-phosphate
• Colorimetric assay
• Glucose-6-phosphate dehydrogenase

A specific colorimetric assay for the determination of glucose-6-phosphate (G6P) was developed. This assay is based on the oxidation of G6P in the presence of glucose-6-phosphate dehydrogenase (G6PD) and nicotinamide adenine dinucleotide phosphate (NADP(+)); the NADPH thereby generated reduces the tetrazolium salt WST-1 [2-(4-indophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H tetrazolium, monosodium salt] to water-soluble yellow-colored formazan with 1-methoxy-5-methylphenazium methylsulfate (1-mPMS) as an electron carrier. The assay is optimized for reaction buffer pH, enzyme/dye concentration, and reaction time course. The limit of detection of the assay is 0.15 mu M (15 pmol/well). The usefulness of the assay is demonstrated by the accurate measurement of the G6P concentration in fetal bovine serum (FBS).

• 出版日期2011-12-15
• 单位NIH