Description

Nori Equine OSM ELISA Kit Summary

Alternative names for OSM: Oncostatin M

Alternative name for equine: Horse

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background: 

Oncostatin M (OSM) is a protein that is encoded by the OSM gene.^[1] OSM is a pleiotropic cytokine that belongs to the interleukin 6 group of cytokines.^[2] Of these cytokines it most closely resembles leukemia inhibitory factor (LIF) in both structure and function.^[2] However, it is as yet poorly defined and is proving important in liver development, haematopoeisis, inflammation and possibly CNS development. It is also associated with bone formation and destruction.^[3] OSM signals through cell surface receptors that contain the protein gp130. The type I receptor is composed of gp130 and LIFR, the type II receptor is composed of gp130 and OSMR.^[4] The role of OSM as an inflammatory mediator was clear as early as 1986.^[5] however, its precise effect on the immune system is far from clear although it can serve as both pro-inflammatory and anti-inflammatory cytokine. OSM is synthesized by stimulated T-cells and monocytes.^[6] The effects of OSM on endothelial cells suggest a pro-inflammatory role for OSM. Endothelial cells possess a large number of OSM receptors.^[7] Stimulation of a primary endothelial culture (HUVEC) with hOSM results in delayed but prolonged upregulation of P-selectin,^[8] which facilitates leukocyte adhesion and rolling, necessary for their extravasation. OSM also promotes the production of IL-6 from these cells.^[7] OSM reduces the degree of joint destruction in an antibody induced model of rheumatoid arthritis.^[9] OSM is a major growth factor for Kaposi’s sarcoma “spindle cells”, which are of endothelial origin.^[10] These cells do not express LIFR but do express OSMR at high levels.^[11] For example, OSM can modulate the expression of IL-6, an important regulator of the host defence system.^[7] OSM can regulate the expression of acute phase proteins. OSM regulates the expression of various protease and protease inhibitors, for example Gelatinase and a1-chymotrypsin inhibitor.

References

1. Rose TM, Bruce AG (1991). Proc. Natl. Acad. Sci. U.S.A. 88 (19): 8641–5.
2. Tanaka M, Miyajima A (2003). Rev. Physiol. Biochem. Pharmacol. 149: 39–52.
3. Walker EC, et al. (2010). J Clin Invest. 120 (2): 582–92.
4. Auguste P, et al. (1997). J. Biol. Chem. 272 (25): 15760–4.
5. Zarling JM, et al. (1986). Proc. Natl. Acad. Sci. U.S.A. 83 (24): 9739–43.
6. Malik N, et al. (1989). Mol. Cell. Biol. 9 (7): 2847–53.
7. Brown TJ, et al. (1991). J. Immunol. 147(7): 2175–80.
8. Yao L, et al. (1996). J. Exp. Med. 184 (1): 81–92.
9. Wallace PM, et al. (1999). J. Immunol. 162 (9): 5547–55.
10. Nair BC, et al. (1992). Science. 255 (5050): 1430–2.
11. Murakami-Mori K, et al. (1995). J. Clin. Invest. 96 (3): 1319–27.

Product Citation