Human IL-6 ELISA Product Overview

The Biomedica human Interleukin-6 ELISA (IL-6) ELISA kit is a sandwich enzyme immunoassay that has been optimized and fully validated for the quantitative determination of human IL-6 in serum and plasma (EDTA, heparin, citrate).  Validation experiments have been performed according to international quality guidelines (ICH/ FDA/ EMEA).  Cell culture supernatant and urine samples are compatible with this ELISA.  The IL-6 ELISA assay recognizes both natural and recombinant human IL-6. The assay employs highly purified epitope mapped antibodies as well as human serum-based standards and controls.

Human IL-6 ELISA Assay Principle

The figure below explains the principle of the human IL-6 sandwich ELISA:

Capture antibody: recombinant anti-human IL-6 antibody
Detection antibody: polyclonal anti-human IL-6 antibody, Biotin-labeled
Target antigen: human IL-6

This kit is a sandwich enzyme immunoassay for the quantitative determination of human IL-6 (Interleukin-6) in human serum, plasma, cell culture supernatants, and urine samples.  In a first step, STD/sample/CTRL are pipetted into the wells, which are pre-coated with the recombinant anti-human IL-6 antibody.  Any soluble IL-6 present in the STD/sample/CTRL binds to the pre-coated anti-IL-6 antibody in the well. After incubation, a washing step is applied where all non-specific unbound material is removed. In a next step, the biotinylated anti-IL-6 antibody (AB) is pipetted into the wells and reacts with the IL-6 present in the sample, forming a sandwich. Next, all unbound antibody is removed during another washing step. In the next step, the conjugate (streptavidin-HRPO) is added and reacts with the biotinylated anti-IL-6 antibody. After another washing step, the substrate (Tetramethylbenzidine; TMB) is pipetted into the wells. The enzyme catalysed colour change of the substrate is directly proportional to the amount of IL-6 present in the sample. This colour change is detectable with a standard microtiter plate ELISA reader. A dose response curve of the absorbance (optical density, OD at 450 nm) versus standard concentration is generated, using the values obtained from the standards. The concentration of soluble IL-6 in the sample is determined directly from the dose response curve.