Principle of Assay
Mouse Lipocalin-2/NGAL ELISA Kit (Small Sample Volume) (A323236) employs the sandwich Enzyme-Linked Immunoassay technique for the quantitative measurement of mouse in serum, plasma, and other biological fluids. This assay is optimized for limited sample availability, requiring only 25 µL of sample per well. An antibody specific for Lipocalin-2/NGAL has been pre-coated onto a 96-well microtiter plate. The standards and test samples are added into the wells and the Lipocalin-2/NGAL present in each sample is bound to the wells by the immobilized antibody. Following incubation, the wells are washed and then incubated with biotinylated anti-Lipocalin-2/NGAL antibody, which binds the captured Lipocalin-2/NGAL present in each well. Following incubation, unbound biotinylated detection antibody is removed by washing, and an HRP-Avidin conjugate is added to the wells and the microtiter plate is incubated. Following incubation and washing, TMB substrate solution is then used to visualize the HRP enzymatic reaction by catalysis to produce a blue-coloured product that changes to yellow after addition of acidic stop solution. The density of yellow is proportional to the amount of Lipocalin-2/NGAL captured in each well. The concentration of Lipocalin-2/NGAL can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
