AffiELISA® Pan 5-HETE Kit: Ultra-High 5-Hydroxyeicosatetraenoic Acid ELISA Detection

The AffiELISA® Pan 5-HETE Kit uses competitive inhibition enzyme immunoassay technique. The provided microtiter plate comes pre-coated with 5-Hydroxyeicosatetraenoic Acid (5-HETE) protein. To perform the assay, standards or samples are added to designated wells on the microtiter plate. Subsequently, a biotin-conjugated antibody specific to 5-Hydroxyeicosatetraenoic Acid (5-HETE) 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 5-Hydroxyeicosatetraenoic Acid (5-HETE) in the samples can be determined.