Interleukin 2 s-Receptor-alpha, Human (IL-2sRa), BioAssay(TM) ELISpot Kit

Intended Use:\nThis 2.5 hour ELISpot kit is developed to detect and visualize of single cells secreting human IL-2sRa.\n\nPrinciple:\nAdd stimulated cells or cells and stimulant to the wells and incubate at 37 degrees C in CO2 incubator for a specified period. Secreted human IL-2sRa binds to antibody coated microtiter plate.\nmicrotiteer plate coated with human IL-2sRa capture Antibody. Cells and unbounded proteins are washed away. Streptavidin-AP or Streptavidin-HRP is added and binds to the biotinylated detection antibody. Unbounded proteins are washed away. Biotinylated detection antibody is added and binds to the secreted human IL-2sRa. Substrate Solution is added. A colored precipitate forms and appears as spots at the sites of human IL-2sRa secreting location. Each individual spot representing an individual human IL-2sRa secreting cell.\n\nBackground:\nThe biological function of IL-2 is obtained by binding to the specific interleukin-2 receptor (IL-2R). The IL-2R consists of three non-covalently linked chains, all of which are type I transmembrane proteins and include the a chain (IL-2Ra, p55), b chain (IL-2Rb, p75), and g chain (IL-2Rg, p65). The a chain is cleaved from the cell surface via nonspecific proteolysis.\n\nIL-2Ra and IL-2Rbg dimers bind to different residues on the IL-2 protein. The IL-2Ra complex displays low affinity and the IL-2Rab complex displays intermediate affinity for IL-2 binding. Both IL-2Ra and IL-2Rab complexes are unable to transduce a signal. The IL-2Rbg complex has intermediate affinity for IL-2 binding and can transduce a signal with a relatively high concentration of IL-2. The IL-2Rabg trimer is the high-affinity receptor for IL-2 and can transduce a signal successfully.\n\nMany cells are capable of expressing IL-2Ra including the antigen-activated T cells and B cells, and approximately 10% of natural killer (NK) cells, leukemia and lymphoma cells. When produced by activated T cells, the a chain is 10-20 folds in excess of the b and g chain. A soluble IL-2Ra can be detected in tissue culture media of IL-2R+ cells and in the serum of experimental animals and humans undergoing an immune response.\n\nThe major biological activities of IL-2R include promoting the proliferative expansion of T cells and NK cells upon activation, promoting the persistence of antigen-selected memory T cells, and promoting homeostasis of the immune system after it has successfully responded to an antigen. However, the biological activity of soluble IL-2Ra is unclear. It has been reported that elevated IL-2 sRa level is accompanied by increased T and B cell activation and immune system activation as observed in rheumatoid arthritis, systemic lupus erythematosis (SLE), some leukemias and lymphomas. Because of its low affinity, IL-2 sRa would be expected to be an inhibitor of IL-2.\n\nKit Components:\nA. Microtiter Plates 1x96wells PVDF-bottom Immunospot plates pre-coated with mouse anti-human IL-2sRa monoclonal antibody. \nB. Positive Control 1x1 vial. Supplied as a lyophilized recombinant human IL-2sRa. Reconstitute with 250 ul Cell Culture Media before use. Use in 1 hour. \nC. Wash Buffer (20x) 1x60ml. Add 1 volume of Wash Buffer (20x) to 19 volume of ddH2O. Use in 1 week. Stored at room temperature. \nD. Human IL-2sRa (Biotin) 1x11ml Biotinylated mouse anti-human IL-2sRa monoclonal antibody Ready to use. \nE. Streptavidin (AP)(100x) 1x120ul. Add 1 volume of Streptavidin (AP)(100x) to 100 volumes of F. (Streptavidin (AP) Diluent) before use. Use in 1 month. Stored at 4 degrees C. \nF. Streptavidin (AP) Diluent 1x11ml. Ready to use.\nG. BCIP/NBT Substrate Solution 1x11ml. Ready to use.\n\nStorage and Stability: \nStore other components at 4C. 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

I7663-28L

Size

96Tests

Applications

ELISA

Reactivities

Hum

References

1. Chilosi, M., et al. (1987) Blood 70:1530.\n2. Fernandez-Botran, R. (1991) FASEB J. 5: 2567.\n3. Harrington, D.S., et al. (1988). Arch. Pathol. Lab Med. 112: 597.\n4. Hatakeyama, M., et al. Science 244, 551-556.\n5. Leonard, W.J., et al. (1994) Immunol Rev. 138: 61.\n6. Leonard, W.J., et al. (1994) Curr. Opin. Immunol. 6: 631.\n7. Minami, Y., et al. (1993). Annu. Rev. Immunol 11:245.\n8. Pizzolo, G., et al. (1987) Br. J. Haematol. 67: 377.\n9. Robb, R. J., et al. J. Exp. Med. 160, 1126-1146.\nS7.5(00) IL-2sRa SL10033E 5\n10. Rubin, L.A., et al. (1985) Hybridoma 4: 91.\n11. Semenzato, G., et al. (1987) Blood 709: 396.\n12. Sharon, M., et al. Science 234, 859-863.\n13. Smith, K.A. (1989) Annu. Rev. Cell Biol. 5:397.\n14. Steis, R.G., et al. (1988). Blood 71: 1304.\n15. Takeshita, T., et al. J. Immunol. 148, 2154-2158.\n16. Taniguchi, T and Y. Minami (1993) Cell 73: 5.\n17. Teshigawara, K., et al. J. Exp. Med. 165, 223-238.\n18. Tsudo, M., et al. Proc. Natl Acad. Sci. USA 83, 9694-9698.\n19. Voss, S.D., et al. (1994) Blood 83: 626.\n20. Wagner, D.K., et al. (1987) J. Clin. Oncol. 5:1262.\n21. Waldmann, T.A. (1991) J. Biol. Chem. 266: 2681.\n22. Waldmann, T.A. (1993) Immunol. Today 14: 264.\n23. Wolf, R.E., et al. (1988) Arthritis Rheum. 31: 729.\n24. Wang, H. M., et al. J. Exp. Med. 166, 1055-1069.