AffiELISA® Sheep NEFA Kit: High Sensitivity Non-Esterified Fatty Acid ELISA Detection

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