RGM-A, ELISA BioAssay(TM) Kit (Repulsive Guidance Molecule A, RGM Domain Family Member A, Rgma)

Mouse Repulsive Guidance Molecule (RGM-A) is a 33kD member of the RGM family of GPI-linked membrane associated proteins (1, 2). It is synthesized as a 454aa prepro-protein that contains a 47aa N-terminal signal sequence and a 27aa C-terminal GPI attachment signal (3, 4). As a result of intramolecular cleavage, mature mouse RGM-A is a disulfide-linked dimer of the 122aa N-terminal and 258 C-terminal segments. The N-terminal segment contains an RGD tripeptide and two potential N-linked glycosylation sites, while the C-terminal segment contains an abbreviated von Willebrand factor domain (3-5). Mouse RGM-A shares 86%, 94%, and 100% aa sequence identity with chicken, human, and rat RGM-A, respectively. It also shares 56-48% aa sequence identity with RGM-B and -C, respectively.\n\nRGM-A is expressed in the developing central nervous system in a pattern that is complementary to the expression of RGM-B (3, 4). It is widely expressed on neuronal and glial cells of the mouse nervous system, particularly in the developing forebrain, the periventricular layers of the cerebrum, the optic tectum/superior colliculus of the midbrain, the retina, and in enteric ganglia of the fetal and adult gut (3-9). Among non-neuronal tissues, RGM-A mRNA is expressed in the gut, heart, lung, liver, skin, kidney, and testes (6, 8, 10).\n\nRGM-A regulates neuronal survival and development through interactions with a receptor complex that includes Neogenin and UNC5H2 (11, 12). The pattern of Neogenin expression overlaps tissue regions that exhibit RGM-A responsiveness (11, 13). RGM-A acts as a nervous system morphogen and as an axon guidance molecule. It promotes neuronal survival, inhibits neurite outgrowth, and induces growth cone collapse (5, 9, 12-15). RGM-A regulates the layer-specific termination of entorhinal complex axons into the hippocampus (16). It also guides retinal ganglion cell axons through the optic fissure to their accurate termination in the optic tectum (5, 17-20). Following tissue damage, RGM-A exerts prosurvival and neuroprotective effects on injured retinal ganglion cells (14). Its upregulation in the vicinity of spinal cord injuries can also hinder neurological regeneration by inhibiting axonal regeneration and synapse formation (7, 21).\n\nSimilarly to other RGM family members, RGM-A also functions as a co-receptor that enhances signaling by BMP-2 and BMP-4 (10). The exact BMP signaling function that is mediated by RGM-A remains to be determined. Recently, RGM-B has been shown to be an important negative regulator of IL-6 expression in immune cells (22). The RGM-A gene has been shown to be associated with experimental inflammation and multiple sclerosis, suggesting that variants of the RGM-A gene can differentially affect immunoregulation and be associated with pro-inflammatory cytokines and antibody responses (23). In colorectal cancer, downregulation of RGM-A correlates with increased tumor cell proliferation and invasion (24).\n\nThe Mouse/Rat RGM-A Immunoassay is a 4.5h solid phase ELISA designed to measure mouse and rat RGM-A in cell culture supernates, tissue homogenates, serum, and plasma. It contains NS0-expressed recombinant mouse RGM-A and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant factor accurately. Results obtained using natural RGM-A showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural RGM-A.\n\nSample Type:\nCell Culture Supernates-Remove particulates by centrifugation and assay immediately or aliquot and store samples at E -20 degrees C. Avoid repeated freeze-thaw cycles.\n\nTissue Homogenates-The preparation of tissue homogenates will vary depending upon tissue type. For this assay, brain tissue from individual mice was removed, rinsed in 1X PBS, and kept on ice. Using a tissue homogenizer, the tissue was homogenized in 1X PBS and frozen at

Prices direct from United States Biological

Quick response times

Exclusive Biosave savings/discounts

SPECIFICATIONS

Catalog Number

R1994-87

Size

1Kit

References

1. Corradini, E. et al. (2009) Cytokine Growth Factor Rev. 20:389. 2. Severyn, C.J. et al. (2009) Biochem. J. 422:393. 3. Schmidtmer, J. and D. Engelkamp (2004) Gene Expr. Patterns 4:105. 4. Niederkofler, V. et al. (2004) J. Neurosci. 24:808. 5. Monnier, P.P. et al. (2002) Nature 419:392. 6. Oldekamp, J. et al. (2004) Gene Expr. Patterns 4:283. 7. Hata, K. et al. (2006) J. Cell Biol. 173:47. 8. Metzger, M. et al. (2005) Dev. Dyn. 234:169. 9. Metzger, M. et al. (2007) J. Neurochem. 103:2665.10. Babitt, J.L. et al. (2005) J. Biol. Chem. 280:29820. 11. Rajagopalan, S. et al. (2004) Nat. Cell Biol. 6:756. 12. Hata, K. et al. (2009) J. Cell Biol. 184:737. 13. Matsunaga, E. et al. (2004) Nat. Cell Biol. 6:749. 14. Koeberle, P.D. et al. (2010) Neuroscience 169:495. 15. Yoshida, J. et al. (2008) Biochem. Biophys. Res. Commun. 372:725. 16. Brinks, H. et al. (2004) J. Neurosci. 24:3862. 17. Tassew, N.G. et al. (2008) Mol. Cell. Neurosci. 37:761. 18. Wilson, N.H. and B. Key (2006) Dev. Biol. 296:485. 19. Schnichels, S. et al. (2007) Gene Expr. Patterns 8:1. 20. Matsunaga, E. et al. (2006) J. Neurosci. 26:6082. 21. Kyoto, A. et al. (2007) Brain Res. 1186:74. 22. Xia, Y. et al. (2011) J. Immunol. 186:1369. 23. Nohra, R. et al. (2010) Genes Immun. 11:279. 24. Li, V.S. et al. (2009) Gastroenterology 137:176.