This Human 12-hydroxyeicosatetraenoic acid (12-HETE) ELISA Kit employs a two-site sandwich ELISA to quantitate 12-HETE in samples. An antibody specific for 12-HETE has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any12-HETE present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for 12-HETE is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of 12-HETE bound in the initial step. The color development is stopped and the intensity of the color is measured.
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