IL-12, Mouse, BioAssay(TM) ELISpot Kit (Interleukin 12, IL-23 p40, CLMF2, IL12B, IL-12B, NKSF2)

Interleukin 12 (IL-12), also known as natural killer cell stimulatory factor (NKSF) or cytotoxic lymphocyte maturation factor (CLMF), is a heterodimeric pleiotropic cytokine made up of a 40kD (p40) subunit and a 35kD (p35) subunit. The IL-12 p40 subunit is shared by IL-23, another heterodimeric cytokine that has biological activities similar to, as well as distinct from, IL-12. IL-12 is produced by macrophages and B cells and has been shown to have multiple effects on T cells and natural killer (NK) cells. While mouse IL-12 is active on both human and mouse cells, human IL-12 is not active on mouse cells.\n\nSample Type:\nWhole Cells\n\nIntended Use:\nThe Mouse IL-12/IL-23 p40 BioAssay(TM) ELISpot assay is designed for the detection of IL-12/IL-23 p40 secreting cells at the single cell level, and it can be used to quantitate the frequency of mouse IL-12/IL-23 p40 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 CD8 T cell responses. \n\nTest Principle:\nBioAssay(TM) ELISpot assays employ the quantitative sandwich ELISA technique. A polyclonal antibody specific for mouse IL-12/IL-23 p40 has been pre-coated onto a PVDF backed microplate. Appropriately stimulated cells are pipetted 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-12/IL-23 p40. After washing away any cells and unbound substances, a biotinylated polyclonal antibody specific for mouse IL-12/IL-23 p40 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-12/IL-23 p40 secreting cell. \n\nKit Components:\nMouse IL-12/IL-23 p40 Microplate, 1x96wells\nDetection Antibody Concentrate, 1x150ul\nStreptavidin-AP Concentrate A, 1x150ul\nDilution Buffer 1, 1x12ml\nDilution Buffer 2, 1x12ml\nWash Buffer Concentrate, 10X concentrated, 1x50ml\nBCIP/NBT Chromogen, 1x12ml\nMouse IL-12/IL-23 p40 Positive Control, lyophilized, 1x vial\n\nStorage and Stability:\nStore powder at 4 degrees C liquid at -20 degrees C. Store other components at 4 degrees C. Stable for 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

145747

Size

1Kit

References

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