This immunoassay kit allows for the specific measurement of rabbit Oncostatin-M, OSM concentrations in cell culture supernates, serum and plasma.
OSM is a cytokine originally isolated from medium conditioned by PMA-treated U-937 histiocytic leukemia cells based on its ability to inhibit growth of A375 melanoma cells. The OSM cDNA encodes a 252 amino acid pre-pro-OSM polypeptide with a 25 residue hydrophobic signal peptide and a hydrophilic C-terminal domain that are proteolytically processed to generate the 196 residue mature form of OSM. Although both mature and pro-OSM are equally active in radio-receptor assays, the mature OSM is 5- to 60-fold more active in growth inhibition assays. Thus, proteolytic processing of the pro-OSM peptide may be important in regulating the in vivo activities of OSM.
OSM is a pleiotropic cytokine that initiates its biological activities by binding to specific cell surface receptors. Recently, gp130, a signal transducing component (β subunit) of the IL-6, LIF and CNTF receptor complexes, was identified as a low-affinity OSM receptor that does not transduce OSM signals. The low affinity LIF receptor (LIF R, a gp130-related protein) has now been identified to be a component of a high-affinity OSM receptor that will transduce OSM signals. Since OSM is also active on cells that do not express LIF R, a specific OSM receptor that does not involve LIF R must also exist. Besides its growth inhibitory activities on A375 melanoma and mouse M1 myeloid leukemic cells, as well as on other solid tumor cells, OSM also has growth stimulatory activities on normal fibroblasts, AIDS-Kaposi’s sarcoma cells, and a erythroleukemia cell line, TF-1. Other OSM-mediated activities reported to date include: stimulation of plasminogen activator activity in cultured bovine aortic endothelial cells; regulation of IL-6 expression in endothelial cells; and stimulation of LDL uptake and up-regulation of cell surface LDL receptors in HepG2 cells.
This assay employs the quantitative sandwich enzyme immunoassay technique. A antibody specific for OSM has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any OSM present is bound by the immobilized antibody. An enzyme-linked antibody specific for OSM is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of OSM bound in the initial step. The color development is stopped and the intensity of the color is measured.