Human IL-7 is 17.4kD protein containing six exons. The gene has a length of ~33kb and maps to chromosome 8q12-q13. Expressed mouse IL-7 is a 25kD single-chain glycoprotein. Human (152 amino acids, 17.4kD) and murine IL-7 (129 amino acids) display a 60 percent sequence homology at the protein level, and both proteins contain three disulfide bonds that are essential for the biological activity of the molecule. The human IL-7 receptor is an integral, highly glycosylated membrane protein of 76kD that is expressed on activated T-cells, which has been designated as CD127.IL-7 is produced predominantly by epithelial cells, especially keratinocytes and thymic epithelial cells, and is a nonredundant cytokine in thymic T-cell development. IL-7 enhances survival and proliferation of T cell precursors in humans and mice and B cell precursors in mice. IL-7 also affects survival and proliferation of mature T cells, but not mature B cells, and is known to be a potent enhancer of T cell function and IFN-y production. IL-7 system deficiency in humans may cause severe combined immunodeficiency (SCID) syndrome.IL-7 is acritical cytokine for normal T and B lymphopoiesis and is a mobilizer ofpluripotent stem cells and myeloid progenitors. IL-7 enhances T cell function and induces cytokine expression in monocytes. Preclinical studies indicate that IL-7 is capable of accelerating murinelymphocyte regeneration following chemotherapy and bone marrowtransplantation, inducing anti-tumor effects in mice, and expandinganti-HIV-specific human T cells. Because of the potent capability of IL-7 in T-lymphopoiesis augmentation, IL-7 may play a role in the immune reconstitution of T-cells during HIV infection, especially in the context of effective antiretroviral treatments. IL-7 is elevated in untreated patients with advanced HIV disease and in those who fail highly active antiretroviral therapy (HAART), but is undetectable in HIV-infected patients who respond to HAART. Immune reconstitution is a critical issue in the treatment of HIV infection, and immune replacement therapy that relies on a variety of regulatory cytokines has been proposed to reconstitute the immune system function in HIV-infected patients. Thus, it is essential that continuedpreclinical and clinical research is performed to evaluate IL-7 as apotential therapy for leukopenia, bone marrow/stem cell transplantation,cancer, and HIV/AIDS. This IL-7 ELISA is a ready-to-use 3.5-hour solid phase immunoassay readily capable of measuring IL-7 levels in serum, plasma, cell culture supernatant, and other biological fluids in the range of 0 to 2000pg/ml. This 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-7 and the various conditions mentioned.Intended Use:This Human Interleukin-7 ELISA Kit is to be used for the in vitro quantitative determination of human interleukin-7 (IL-7) concentrations in serum, plasma, cell culture supernatant, and other biological fluids. Sensitivity:<1pg/mlRange:1.5-500pg/mlTest Principle:This IL-7 enzyme linked immunosorbent assay (ELISA) 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-7. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for IL-7 and incubated. If present, IL-7 will bind and become immobilized by the antibody pre-coated on the wells and then become “sandwiched” by biotin conjugate. The microtiter plate wells are thoroughly washed to remove unbound IL- 7 and other components of the sample. In order to quantitatively determine the amount of IL-7 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-7, 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 450nm ± 2nm.In order to measure the concentration of IL-7 in the samples, this 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 provided standard 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-7 concentration (pg/ml). The concentration of IL-7 in the samples is then determined by comparing the O.D. of the samples to the standard curve.Kit Components:IL-7 MICROTITER PLATE BIOTIN CONJUGATE AVIDIN CONJUGATE IL-7 STANDARD CALIBRATOR DILUENT I CALIBRATOR DILUENT II WASH BUFFER (20X/96 wells, 30X/192 wells) SUBSTRATE A SUBSTRATE B STOP SOLUTIONStorage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.