General Estradiol ELISA Kit

The Antibodies.com General Estradiol ELISA Kit is a competitive Enzyme-Linked Immunosorbent Assay (cELISA) for the in vitro quantitative determination of Estradiol in serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids. The assay uses a pre-coated microplate format with colorimetric detection read at 450 nm on a microplate reader.

The competitive enzyme immunoassay technique involves Estradiol in samples or standards competing with immobilized Estradiol antigen for binding sites on a Biotinylated Anti-Estradiol Antibody. After incubation and washing, an HRP-Avidin conjugate is added, and TMB substrate is used to visualize the HRP enzymatic reaction, producing a blue colour that changes to yellow upon addition of acidic stop solution. The density of yellow is inversely proportional to the amount of Estradiol present, and concentration is calculated by reference to a standard curve.

Performance specifications include a product range of 0.156–10 ng/ml, a sensitivity of <0.062 ng/ml, and an assay time of 2h 30m. Recovery validation showed serum averaging 91% (range 80%–102%; n=5), EDTA plasma averaging 90% (range 81%–99%; n=5), and heparin plasma averaging 84% (range 80%–89%; n=5). Linearity at 1:2, 1:4, 1:8, and 1:16 dilutions was reported for serum (87–91%, 87–107%, 74–101%, 92–97%; n=5), EDTA plasma (90–105%, 84–101%, 90–101%, 79–108%; n=5), and heparin plasma (84–95%, 92–105%, 82–105%, 89–91%; n=5). Kit components should be stored at +4°C or -20°C as instructed, and the kit should not be used past its expiration date.