IL-11 ELISA KIT
INTENDED USE
This Human IL-11 ELISA Kit is to be used for the in vitro quantitative determination of human Interleukin 11 (IL-11) concentrations in serum, plasma, and cell culture supernatant. This IL-11 ELISA Kit is intended for LABORATORY RESEARCH USE ONLY and is not for use in diagnostic or therapeutic procedures.
INTRODUCTION
Interleukin-11, also known as adipogenesis inhibitory factor (AGIF) and oprelvekin, is a member of the GP130 family of cytokines. These cytokines drive the assembly of multi-subunit receptor complexes containing at least one GP130. IL-11 exerts biological activity through IL-11 receptor by utilizing the gp130 molecule as the signaling component of its receptor. Interleukin 11 (IL-11) is a highly conserved precursor protein of 19kDa with 199 amino acids. The first 21 amino acids are a typical leader sequence for secreted proteins. When the 178 amino acid protein is released from the cell, the first 21 amino acids are removed. IL-11 is thermally stable and contains no cysteine residues. Many cell and tissue types throughout the body, such as CNS, thymus, lung, bone, connective tissues, uterus, skin, and testis, can produce IL-11 depending on the local environment of the cell or tissue. IL11 promotes primary and secondary immune responses in vitro and in vivo and modulates antigen-specific antibody reactions. IL-11 has activity in bone metabolism and activity in protection and restoration of the gastrointestinal mucosa. Human endometrial stromal cells produce biologically active IL-11, which promotes progesterone-induced decidualization. Therefore, the IL-11 has both paracrine and autocrine actions on human endometrial stromal cells and plays an important role in preparing the human endometrium for implantation. Reports indicate that abnormal IL-11 levels in serum or plasma is related to immune thrombocytopenia, rheumatoid arthritis, and urogenital infection. Immunoreactive IL-11 has also been linked to aseptic loosening of total hip replacement implants. The production of IL-11 is elevated in lesional skin organ cultures of patients with active plaque-type psoriasis as compared with non-lesional and normal skin.
The human IL-11 genes contain five exons, each having a length of 7 kDa, and are mapped to chromosome 19q13. 3-13. 4. Primate and human IL-11 sequences show 97 percent homology. The human IL-11 receptor gene maps to chromosome 9p13.
PRINCIPLE OF THE ASSAY
IL-11 ELISA Kit
IL-11 ELISA Kit is to be used for the in vitro quantitative determination of human Interleukin 11 (IL-11) concentrations in serum, plasma, and cell culture supernatant. This IL-11 ELISA Kit is intended for LABORATORY RESEARCH USE ONLY and is not for use in diagnostic or therapeutic procedures. This IL-11 ELISA Kit is a ready-to-use 3.5-hour solid phase immunoassay capable of measuring IL-11 levels in serum, plasma, cell culture supernatant in the range of 0 to 1600 pg/mL. This IL-11 ELISA Kit has been shown to have a specific reaction with IL-11 without any cross reactivity with various other cytokine super-family proteins. This IL-11 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-11. Standards or samples are then added to the appropriate microtiter plate wells and incubated. After washing to remove unbound IL-11 and other components of the sample, biotin-conjugated polyclonal antibody specific to IL-11 is added and incubated. If present, IL-11 will bind and become immobilized by the antibody pre-coated on the wells and then become “sandwiched” by the biotin conjugate. In order to quantitatively determine the amount of IL-11 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-11, biotin-conjugated antibody, and enzyme-conjugated Avidin will exhibit a change in colour. 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.
In order to measure the concentration of IL-11 in the samples, this IL-11 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-11 concentration (pg/mL). The concentration of IL-11 in the samples is then determined by comparing the O.D. of the samples to the standard curve.
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