Interleukin 2, Primate, BioAssay(TM) ELISpot Kit (IL-2)

Interleukin 2 (IL-2) was initially identified as a T cell growth factor that is produced by T cells following activation by mitogens or antigens. Since then, it has also been shown to stimulate the growth and differentiation of B cells, natural killer (NK) cells, lymphocyte activated killer (LAK) cells, monocytes/macrophages and oligodendrocytes. At the amino acid sequence level, there is approximately 60% - 90% similarity between species. Mature human IL-2 shows 65%, 67%, 72%, 78%, and 64% aa identity to mouse, rat, pig, cat, and cow IL-2, respectively.\n\nIntended Use:\nFor the quantitative determination of the frequency of cells releasing primate IL-2.\n\nTest Principle:\nThe enzyme-linked immunospot (ELISpot) assay was originally developed for the detection of\nindividual B cells secreting antigen-specific antibodies (21, 22). This method has since been\nadapted for the detection of individual cells secreting specific cytokines or other antigens\n(23, 24). ELISpot assays employ the quantitative sandwich enzyme-linked immunosorbent\nassay (ELISA) technique. A monoclonal antibody specific for primate IL-2 has been pre-coated\nonto a PVDF (polyvinylidene difluoride)-backed microplate. Appropriately stimulated cells are\npipetted into the wells and the microplate is placed into a humidified 37 degrees C CO2 incubator for a specified period of time. During this incubation period, the immobilized antibody in the immediate vicinity of the secreting cells bind secreted IL-2. After washing away any cells and unbound substances, a biotinylated polyclonal antibody specific for primate 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\nand 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\nsystem or manually using a stereomicroscope.\n\nKit Components:\n1. Precoated Microplate, 1x96 wells\n2. Detection Antibody Concentrate, 1x150ul\n3. Streptavidin-AP Concentrate A, 1x150ul\n4. Dilution Buffer 1, 1x12ml\n5. DIlution Buffer 2, 1x12ml\n6. Wash Buffer Concentrate, 1x50ml\n7. BCIP/NBT Chromogen, 1x12ml\n8. Primate IL-2 Positive Control, 1x1vial\n\nStorage and Stability:\nStore components at 4 degrees 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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SPECIFICATIONS

Catalog Number

167669

Size

96Tests

References

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