More info
Assay Range | 15.6 - 1,000 pg/mL |
Sensitivity | 10.0 pg/mL |
Size | 96T |
Storage | Store at 2 - 8ºC. Keep reconstituted standard and detection Ab at -20 ºC |
Assay Principle | Sandwich ELISA |
Sample Volume | 100 µL final volume, dilution factor varies on samples |
Detection Method | Chromogenic |
Kit Components
1. Recombinant Mouse IL-12(p40) standard: 2 vials
2. One 96-well plate coated with anti- Mouse IL-12(p40) Ab
3. Sample diluent buffer: 12 mL — 2
4. Detection antibody: 130 µL, dilution 1:100
5. Streptavidin-HRP: 130 µL, dilution 1:100
6. Antibody diluent buffer: 12 mL x1
7. Streptavidin-HRP diluent buffer: 12 mL x1
8. TMB developing agent: 10 mL x1
9. Stop solution: 10 mL x1
10. Washing solution (20x): 25 mL x1
Background
Interleukin 12 (IL-12), also known as NKSF or p70, is a 70-75 kDa heterodimeric glycoprotein belonging to the IL-12 family. IL-12 is encoded by two separate genes, IL-12A and IL-12B, in human. IL-12 is composed of two disulfide-linked subunits, p35 and p40. The mature p35 subunit is synthesized as a 197 amino acid (aa) containing seven cysteines and one potential N-linked glycosylation site. Mature human p35 shares 58% aa identity with mouse and rat p35. Mature human p40 is a 306 aa protein with 11 cysteines and three potential N-linked glycosylation sites. Mature human p40 is 66% aa identical to mouse and rat p40. IL-12 is produced in macrophages and dendritic cells, monocytes, Langerhans cells, neutrophils, keratinocytes, plasmacytoid dendritic cells, and microglia, etc. p35 mRNA is expressed by various cells and tissues, however, p35 have not been detected in culture supernatants of cells expressing only p35 or both p35 and p40 mRNAs. In cells expressing both p35 and p40 mRNAs, p40 mRNA is expressed at a higher level and free p40 is secreted together with heterodimeric IL-12 p70. Most of the free p40 secreted by the various human cell lines examined have been found to exist as monomers. The p40 and p35 by themselves do not have IL-12 activity, but the homodimer of p40 is capable of binding the IL-12 receptor and is an IL-12 antagonist.
Human IL-12 receptor is composed of at least two type I transmembrane glycoproteins, Rβ1 and Rβ2. Rβ1 is the principal binding protein for the p40 , while Rβ2 might be the principal signal transduction component and serve as an attachment point for a disulfide-linked p35-p40 heterodimer. IL-12 plays an important role in regulating the activities of natural killer cells and T lymphocytes. It enhances cytotoxic activity and induces IFN-γ production in NK cells, T cells and dendritic epidermal T cells.