AffiELISA® Human MTX Kit: Ultra-High Methotrexate ELISA Detection

The AffiELISA® Human MTX Kit uses competitive inhibition enzyme immunoassay technique. The provided microtiter plate comes pre-coated with Methotrexate (MTX) protein. To perform the assay, standards or samples are added to designated wells on the microtiter plate. Subsequently, a biotin-conjugated antibody specific to Methotrexate (MTX) 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 Methotrexate (MTX) in the samples can be determined.