Human HbA1c (glycosylated hemoglobin A1c) ELISA kit is based on a sandwich enzyme-linked immunosorbent assay (ELISA) technology.

A 96-well plate is coated with a specific antibody against the protein of interest (HbA1c), which will bind the protein and this is detected by a biotin conjugated antibody against the protein of interest. The biotin moiety is subsequently detected, following washing, by the addition of streptavidin coupled horseradish peroxidase (HRP). The supplied TMB substrates were used to visualise HRP enzymatic reaction. TMB was catalysed by HRP to produce a blue colour product that changed into yellow after adding acidic stop solution. The density of yellow is directly proportional to the concentration of protein of interest captured on the plate.