AffiELISA® Pan 25-OH-D Kit: High Sensitivity 25 Hydroxy Vitamin D ELISA Detection

The AffiELISA® Pan 25-OH-D Kit uses competitive inhibition enzyme immunoassay technique. The provided microtiter plate comes pre-coated with 25 Hydroxy Vitamin D (25-OH-D) protein. To perform the assay, standards or samples are added to designated wells on the microtiter plate. Subsequently, a biotin-conjugated antibody specific to 25 Hydroxy Vitamin D (25-OH-D) is introduced. Avidin conjugated to Horseradish Peroxidase (HRP) is then added to each well, followed by an incubation period. Afterward, the TMB substrate solution is added, initiating an enzyme-substrate reaction. To conclude the reaction, a sulphuric acid solution is introduced, causing a color change that can be measured spectrophotometrically at a wavelength of 450nm ± 10nm. By comparing the optical density (OD) of the samples to the standard curve, the concentration of 25 Hydroxy Vitamin D (25-OH-D) in the samples can be determined.