IL-2, BioAssay ELISpot Kit (Bovine) (Interleukin 2)-IL-2, BioAssay ELISpot Kit (Bovine) (Interleukin 2)代理/參數(shù)/廠家-細胞分析試劑盒-艾美捷科技有限公司

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中文名稱: IL-2, BioAssay ELISpot Kit (Bovine) (Interleukin 2)

英文名字: IL-2, BioAssay ELISpot Kit (Bovine) (Interleukin 2)

供應商: USBiological

產品貨號: 145749

產品報價: ￥1/1Kit

產品說明書: 點擊查看

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產品描述: Bovine Interleukin 2 (IL-2) is a 15kD, a-helical, single chain, potentially glycosylated polypeptide that has potent stimulatory activity for antigen-activated T cells (1-5). The molecule is synthesized as a 155 amino acid (aa) precursor that contains a 20aa signal peptide plus a 135aa mature segment that is possibly O-glycosylated (4, 5). The mature region has multiple a-helices and one intrachain disulfide bond. Mature bovine IL-2 is 64%, 60%, 49%, 50%, 72%, 63%, and 67% identical to mature human, canine, mouse, rat, porcine, equine, and feline IL-2, respectively. Mammalian cells known to express IL-2 include CD4+ and CD8+ T cells, visceral smooth muscle cells, eosinophils, gd T cells, B cells, and dendritic cells. The receptor for IL-2 is complex and consists of three distinct subunits in varying combinations (6, 7). Two of these are ligand-binding and are termed IL-2 Ra and IL-2 Rb. IL-2Ra is 55kD and binds IL-2 with low affinity. IL-2 Rb is 75kD and binds IL-2 with intermediate affinity. Signal transduction is performed by both IL-2 Rb and a 64kD common gamma chain (gc). This signal transducing common gamma chain does not bind IL-2 but does heterodimerize with IL-2 Rb to form a functional IL-2 receptor. The complex heterotrimeric a-b-gc receptor may arise from IL-2 binding to preformed Ra-Rb complexes (8). Functionally, IL-2 is best known for its autocrine and paracrine activity on T cells. It drives resting T cells into active G1, inducing IL-2 and IL-2 Ra synthesis and cell proliferation (7). It also promotes Fas-induced death of na?ve CD4+ T cells, while having minimal effect on activated CD4+ memory lymphocytes. Finally, IL-2 seems to play a central role in the expansion and maintenance of CD4+ CD25+ regulatory T cells. Thus, IL-2 may be a key cytokine in the natural suppression of autoimmunity (9, 10).Intended Use:IL-2, Bovine BioAssay ELISpot Kit is designed for the detection of IL-2 secreting cells at the single cell level, and it can be used to quantitate the frequency of bovine IL-2 secreting cells. ELISpot assays are well suited for monitoring immune responses to various stimuli, treatments and therapies, and they have been used for the quantitation of antigen-specific CD4 and/or CD8T cell responses. Other methods for the assessment of antigen-specific T cell responses, such as the chromium release assay with quantitation by limiting dilution, are tedious, and require previous in vitro expansion of T cells for several days. These assays typically are not suitable for measuring infrequent T cell responses that occur at less than 1 in 1000. ELISpot assays are highly reproducible and sensitive and can be used to measure responses with frequencies well below 1 in 100,000. ELISpot assays do not require prior in vitro expansion of T cells, and they are suitable for high-throughput analysis using only small volumes of primary cells. As such, ELISpot assays are useful tools for research in areas as diverse as antigen recognition, vaccine development, and the monitoring of various clinical trials.Test Principle:The enzyme-linked immunospot (ELISpot) assay was originally developed for the detection of individual B cells secreting antigen-specific antibodies (11, 12). This method has since been adapted for the detection of individual cells secreting specific cytokines or other antigens (13, 14). ELISpot assays employ the quantitative sandwich enzyme-linked immunosorbent assay (ELISA) technique. A polyclonal antibody specific for bovine IL-2 has been pre-coated onto a PVDF (polyvinylidene difluoride)-backed microplate. Appropriately stimulated cells are pipetted into the wells and the microplate is placed into a humidified 37° C CO2 incubator for a specified period of time. During this incubation period, the immobilized antibody in the immediate vicinity of the secreting cells binds secreted IL-2. After washing away any cells and unbound substances, a biotinylated polyclonal antibody specific for bovine IL-2 is added to the wells. Following a wash to remove any unbound biotinylated antibody, alkaline-phosphatase conjugated to streptavidin is added. Unbound enzyme is subsequently removed by washing and a substrate solution (BCIP/NBT) is added. A blue-black colored precipitate forms and appears as spots at the sites of cytokine localization, with each individual spot representing an individual IL-2 secreting cell. The spots can be counted with an automated ELISpot reader system or manually using a stereomicroscope.Kit Components:Precoated Bovine IL-2 Microplate, 1x96wellsDetection Antibody Concentrate, 1x150ulStreptavidin-AP Concentrate A, 1x50ulDilution Buffer 1, 1x12mlDilution Buffer 2, 1x12mlWash Buffer Concentrate, 1x50mlBCIP/NBT Chromogen, 1x12mlBovine IL-2 Positive Control, 1x1vialStorage and Stability:Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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保存建議: 建議收到IL-2, BioAssay ELISpot Kit (Bovine) (Interleukin 2)產品后將其置于-20°C保存。

其他: Usbiological公司是美國著名的抗體和生化試劑供應商，生產世界上種類最多的抗體，用于Western Blot、免疫沉淀、免疫熒光、免疫組化和流式細胞術等多種檢測方法。Usbiological公司現(xiàn)已擁有超過50,19234種抗體、抗原和生化產品，為科研用戶提供了諸多超值選擇。