Human Interleukin-1 beta ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com designed for the in vitro quantitative determination of human Interleukin-1 beta. The kit applies a quantitative sandwich immunoassay technique, using a microplate pre-coated with a monoclonal antibody specific for IL-1β, and is intended for use with serum, plasma, cell culture supernatant and other biological fluids.

Standards or samples are incubated with a biotin-conjugated monoclonal antibody preparation specific for IL-1β, and any IL-1β present becomes sandwiched between the immobilised capture antibody on the well and the biotin conjugate. After thorough washing to remove unbound components, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each well, followed by a TMB (3,3',5,5' tetramethyl-benzidine) substrate solution. 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 on a colorimetric microplate reader. The assay can detect both natural and recombinant human IL-1β and is reported to show no cross-reactivity with various other cytokines super-family proteins. Two calibration diluents are supplied – Calibrator Diluent I for serum/plasma testing and Calibrator Diluent II for cell culture supernatant testing – to enable construction of a standard curve of Optical Density versus IL-1β concentration.

The reported product range is 2-400 pg/ml with a sensitivity of < 2 pg/ml. The kit is shipped and stored at 4°C and the total assay time is approximately 3 hours 30 minutes.