Description
This Biotin (Serum/Plasma/Urine) ELISA Assay is a competitive ELISA for the determination of biotin in human serum. Samples, standards and controls are pipetted into the wells (pre-coated with streptavidine) and incubated. After a washing step, conjugate (enzyme-labelled biotin) is added and competes against the biotin in the samples, standards and controls for streptavidin on the microtiter plate. Unbound enzyme-labelled biotin is washed away and the enzyme substrate TMB is added, resulting in a colour reaction. Finally, the reaction is terminated by an acidic stop solution causing a colour change from blue to yellow. The color intensity is inversely proportional to the biotin concentration. A dose response curve of the absorbance unit (optical density, OD at 450 nm) vs. concentration is generated using the values obtained from the standards. Biotin present in the samples is determined from this curve