Description
This Human ApoAI and ApoB Duplex ELISA Kit employs the quantitative sandwich enzyme immunoassay technique for the detection and quantitation of both human ApoAI and ApoB (ApoB-100/48) in the same sample of plasma, serum or other biological fluids. Antibodies specific for ApoAI and ApoB has been pre-coated onto a microtiter plate. Standards or samples are pipetted into the wells and any ApoAI and ApoB present are bound by the immobilized antibodies. After washing away any unbound substances, a biotin-conjugated antibody specific for ApoAI and an anti-ApoB antibody are added to each well and incubate. Following a washing to remove unbound substances, streptavidin conjugated to Alkaline Phosphatase (AP) and a Horseradish Peroxidase conjugated secondary antibody reacts to anti-ApoB antibody are added to each microplate well and incubated. After washing away any unbound antibody-enzyme reagent, an AP substrate solution is added to the wells and color develops in proportion to the amount of ApoAI bound in the initial step. The intensity of the color developed from AP substrate is measured at a wavelength of 405nm. After wash away the AP substrate, a HRP substrate solution (TMB) is added to the wells and color develops in proportion to the amount of ApoB bound in the initial step. The color development is stopped by the addition of acid and the intensity of the color is measured at a wavelength of 450nm. The concentration of ApoAI and ApoB in the sample is then determined by comparing the O.D of samples to the standard curve