Human Interleukin-17A ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) supplied by Antibodies.com, designed for the in vitro quantitative determination of human Interleukin-17A in serum, plasma, cell culture supernatant, and other biological fluids. The kit applies a quantitative sandwich immunoassay technique in which the microtiter plate has been pre-coated with a monoclonal antibody specific to IL-17A. Standards or samples are added to the appropriate wells, followed by a biotin-conjugated antibody preparation specific for IL-17A, which is incubated to allow binding. After thorough washing to remove unbound IL-17A and other components, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each well. The wells are washed again to remove all unbound HRP-conjugated Avidin, and a TMB (3,3'5, 5' tetramethyl-benzidine) substrate solution is added. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution, and the colour change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm.

Two calibration diluents are included with the kit: Calibrator Diluent I for serum/plasma testing and Calibrator Diluent II for cell culture supernatant testing. The provided standard is diluted 2-fold with the appropriate Calibrator Diluent and assayed at the same time as the samples, allowing the operator to produce a standard curve of Optical Density (O.D) versus IL-17A concentration (pg/mL) from which sample concentrations are then determined.

The assay recognises both natural and recombinant human IL-17A and has shown no cross-reactivity with other cytokines tested. Reported specifications include a product range of 5-1000 pg/ml, a sensitivity of < 5 pg/ml, and an assay time of 3h 30m. The platform is a pre-coated microplate (12 x 8 well strips), read on a colorimetric microplate reader, and the kit is shipped and stored at 4°C.