PromoCell’s 1,5-Anhydroglucitol Assay Kit is a high-throughput adaptable microplate-based assay that enables rapid quantification of 1,5-AG levels in biological fluids. The assay consists of two steps: in the first step, the sample is pretreated to remove endogenous glucose that may interfere with the 1,5-AG detection and 1,5-AG is converted to 1,5-AG-6-phosphate. In the second step, 1,5-AG-6-phosphate is enzymatically metabolized, generating NADPH, which subsequently reacts with a colorless probe to form a strongly colored product measured at OD = 460 nm. The absorbance is proportional to the concentration of 1,5-AG present in the samples. The assay is not affected by galactose, fructose or other physiologically common sugars and has a limit of detection of 0.11 nmoles of 1,5-AG per well, with a reliable linear range from 0.2 – 5 nmole per well (corresponding to a 20-500 μM concentration in a 10 μl sample volume).
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