IL-10 ELISA KIT
INTENDED USE
This Human IL-10 ELISA Kit is to be used for the in vitro quantitative determination of human interleukin 10 (IL-10) concentrations in serum, plasma, cell culture supernatant, and other biological fluids. This IL-10 ELISA Kit is intended FOR LABORATORY RESEARCH USE ONLY and is not to be used in diagnostic or therapeutic procedures.
INTRODUCTION
The mature human IL-10 is a protein with 160 amino acids and the functional IL-10 is a homodimer. IL-10 is expressed by a variety of cells, including mouse T cells (TH1, TH2, and Tr1 subsets), B cells derived from peripheral blood, tonsils or spleen, Epstein–Barr virus (EBV)-transformed B cell lines, Burkitt’s lymphoma, AIDS lymphomas, monocytes, placental trophoblasts, bronchial epithelial cells, and certain tumor cells including melanomas and carcinomas of various origin.
IL-10 plays an important role in inflammatory and immune responses. The biological activities of IL-10 include both immunosuppressive and immuno-stimulatory effects. For example, IL-10 can suppress the production of pro-inflammatory cytokines by monocytes and neutrophils, and down regulates the expression of activating and co-stimulatory molecules on monocytes and dendritic cells. IL-10 also can improve the growth of B cells and mast cells, and inhibit or enhance the activities of T cells depending on their activation conditions. The different polymorphisms in the IL-10 gene promoter have been demonstrated the association with both SIDS (sudden infant death syndrome) and sudden unexpected death due to infection. Based on the functions of IL-10, it has been suggested that an IL-10 antagonist can be applied to boost anti-viral immunity against viruses such as EBV and that the IL-10 molecule itself may be used as an anti-inflammatory reagent.
PRINCIPLE OF THE ASSAY
IL-10 ELISA Kit
IL-10 ELISA Kit is to be used for the in vitro quantitative determination of human interleukin 10 (IL-10) concentrations in serum, plasma, cell culture supernatant, and other biological fluids. This IL-10 ELISA Kit is intended FOR LABORATORY RESEARCH USE ONLY and is not to be used in diagnostic or therapeutic procedures. This IL-10 ELISA Kit is a ready-to-use 4.5-hour solid phase immunoassay capable of measuring IL-10 levels in serum, plasma, cell culture supernatant, and other biological fluids in the range of 0 to 1600 pg/mL. This IL-10 ELISA Kit assay has shown no cross-reactivity with other cytokines tested, and is expected to be used effectively for further investigations into the relationship between IL-10 and the various conditions mentioned.
This IL-10 ELISA Kit applies a technique called a quantitative sandwich immunoassay. The microtiter plate provided in this kit has been pre-coated with a monoclonal antibody specific to IL-10. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for IL-10 and incubated. IL-10 if present, will bind and become immobilized by the antibody pre-coated on the wells and then will become “sandwiched” by biotin conjugate. The microtiter plate wells are thoroughly washed to remove unbound IL-10 and other components of the sample. In order to quantitatively determine the amount of IL-10 present in the sample, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Avidin is a tetramer containing four identical subunits, each having a high affinity-binding site for biotin. The wells are thoroughly washed to remove all unbound HRP-conjugated Avidin and a TMB (3,3'5, 5' tetramethyl-benzidine) substrate solution is added to each well. The enzyme (HRP) and substrate are allowed to react over a short incubation period. Only those wells that contain IL-10, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm.
In order to measure the concentration of IL-10 within the samples, this IL-10 ELISA Kit includes two calibration diluents (Calibrator Diluent I for serum/plasma testing and Calibrator Diluent II for cell culture supernatant testing.) According to the testing system, the standard provided is diluted (2-fold) with the appropriate Calibrator Diluent and assayed at the same time as the samples. This allows the operator to produce a standard curve of Optical Density (O.D) versus IL-10 concentration (pg/mL). The concentration of IL-10 in the samples is then determined by comparing the O.D. of the samples to the standard curve.
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