Human Interleukin-1 alpha ELISA Kit

From Antibodies.com, this sandwich Enzyme-Linked Immunosorbent Assay (sELISA) is designed for the in vitro quantitative determination of human Interleukin-1 alpha in serum, plasma and cell culture supernatant. The assay applies a quantitative sandwich immunoassay technique using a microtiter plate pre-coated with a monoclonal antibody specific to IL-1a. Standards or samples are added to the appropriate wells and incubated. After washing to remove unbound material, biotin-conjugated polyclonal antibody specific to IL-1a is added, followed by Avidin conjugated to Horseradish Peroxidase (HRP). A TMB (3,3'5,5' tetramethyl-benzidine) substrate solution is then added, the enzyme-substrate reaction is terminated by a sulphuric acid solution, and the colour change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm.

The kit shows no cross-reactivity with various other cytokines super-family proteins and can detect both natural and recombinant human IL-1a. Two calibration diluents are included: Calibrator Diluent I for serum/plasma testing and Calibrator Diluent II for cell culture supernatant testing. The standard provided is diluted 2-fold with the appropriate Calibrator Diluent and assayed alongside the samples to produce a standard curve of Optical Density (O.D.) versus IL-1a concentration in pg/mL.

The product range is 5-500 pg/ml with a sensitivity of < 5 pg/ml. The pre-coated microplate is configured as 12 x 8 well strips and the total assay time is 3h 30m. The kit is shipped and stored at 4°C for reading on a colorimetric microplate reader.