Prolactin ELISA

The Calbiotech Mouse/Rat Prolactin kit is based on a solid phase sandwich ELISA. The samples and diluted biotinylated anti-prolactin antibodies are added to the wells coated with polyclonal antibody to prolactin. Prolactin in samples binds to the anti-prolactin polyclonal antibody on the well and the biotinylated antiprolactin antibody binds to prolactin. Unbound protein and biotin conjugate are washed off by wash buffer. Diluted horseradish peroxidase conjugated streptavidin is added to each wells and the streptavidin is allowed to bind to the biotin in the complex. Unbound enzyme conjugate is washed off. Upon the addition of the substrate, the intensity of color is proportional to the concentration of prolactin in the samples. A standard curve is prepared relating color intensity to the concentration of the prolactin.