Description

The Human Interleukin-6 (IL-6, IL6) ELISA research-use-only kit is to be used for the quantitative determination of IL-6 in samples (see sample types indicated) using 96-well plates and a microplate reader. The assay will recognize both natural and recombinant forms of this target.

Performance characteristics

• Sensitivity: <2>
• Standard curve range: 7.8–500 pg/mL
• Sample type(s): serum, plasma, cell culture supernatant
• Specificity: natural and recombinant human IL-6
• Cross-reactivity: see kit manual for cross-species and/or cross-target reactivity
• Sample volume: 100 μL
• Total assay time: 3 hours

Rigorous validation

Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.

Principle of the method

The human IL-6 solid-phase sandwich ELISA (enzyme-linked immunosorbent assay) is designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody has been pre-coated in the wells of the supplied microplate. Samples, standards, or controls are then added into these wells and bind to the immobilized (capture) antibody. The sandwich is formed by the addition of the second (detector) antibody, binding to the target on a different epitope from the capture antibody. A conjugated enzyme has been incorporated into the assay. After incubation and washing steps to rid the microplate of unbound substances, a substrate solution is added that reacts with the enzyme-antibody-target complex to produce measurable signal. The intensity of this signal is directly proportional to the concentration of target present in the original specimen.

Target information

The Human IL-6 kit is a solid phase sandwich Enzyme-Linked Immunosorbent Assay (ELISA). A monoclonal antibody specific for human IL-6 has been coated onto the wells of the microtiter strips provided. Samples, including standards of known human IL-6 content, control specimens, and unknowns, are pipetted into these wells, followed by the addition of a biotinylated monoclonal second antibody. During the first incubation, the human IL-6 antigen binds simultaneously to the immobilized (capture) antibody on one site, and to the solution phase biotinylated antibody on a second site. After removal of excess second antibody, Streptavidin-Peroxidase (enzyme) is added. This binds to the biotinylated antibody to complete the four-member sandwich. After a second incubation and washing to remove all the unbound enzyme, a substrate solution is added, which is acted upon by the bound enzyme to produce color. The intensity of this colored product is directly proportional to the concentration of human IL-6 present in the original specimen.

Related links

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Learn more about other immunoassays