Antigen: purified enzyme from mouse macrophages (RAW 264.7) cells · Host: rabbit · Application(s): ICC, IP, and WB · Nitric Oxide Synthase (NOS) catalyzes the biosynthesis of nitric oxide from L-arginine. Constitutively expressed NOS is found in brain (nNOS) and endothelial cells (eNOS). iNOS is a soluble enzyme found in a variety of tissues including macrophages, hepatocytes, vascular smooth muscle cells, and chondrocytes.^1,2 iNOS expression is increased by a variety of factors including LPS, IFN-γ, Il-1β, and TNF-α, whereas expression is deceased by dexamethasone.^3,4,5,6,7 The enzyme has been cloned from several species including mouse, rat, and human with homology of at least 80% between these species.² The calculated molecular weight of the protein from the deduced amino acid sequence is 130,000 - 131,000.¹

1 Knowles, R.G., and Moncada, S. Nitric oxide synthases in mammals. Biochem J 298 249-258 (1994).

2 Michel, T., Xie, Q., and Nathan, C. Molecular biological analysis of nitric oxide synthases. 161-175 (1996).

3 Lorsbach, R.B., Murphy, W.J., Lowenstein, C.J., et al. Expression of the nitric oxide synthase gene in mouse macrophages activated for tumor cell killing. Molecular basis for the synergy between interferon-γ and lipopolysaccharide. J Biol Chem 268 1908-1913 (1993).

4 Balligand, J., Ungureanu-Longrois, D., Simmons, W.W., et al. Cytokine-inducible nitric oxide synthase (iNOS) expression in cardiac myocytes. Characterization and regulation of iNOS expression and detection of iNOS activity in single cardiac cyocytes in vivo. J Biol Chem 269 27580-27588 (1994).

5 Schmidt, H.H.H.W., Warner, T.D., Nakane, M., et al. Regulation and subcellular location of nitrogen oxide synthases in RAW264.7 macrophages. Mol Pharmacol 41 615-624 (1992).

6 Ogura, T., and Esumi, H. Nitric oxide synthase expression in human neuroblastoma cell line induced by cytokines. Mol Neurosci 7 853-856 (1996).

7 Walker, G., Pfeilschifter, J., and Kunz, D. Mechanism of suppression of inducible nitric-oxide synthase (iNOS) expression in interferon (IFN)-γ-stimulated RAW 264.7 cells by dexamethasone. Evidence for glucocorticoid-induced degradation of iNOS protein by calpain as a key step in post-transcriptional regulation. J Biol Chem 271 16679-16687 (1996).

Knowles, R.G., and Moncada, S. Nitric oxide synthases in mammals. Biochem J 298 249-258 (1994).

Lorsbach, R.B., Murphy, W.J., Lowenstein, C.J., et al. Expression of the nitric oxide synthase gene in mouse macrophages activated for tumor cell killing. Molecular basis for the synergy between interferon-γ and lipopolysaccharide. J Biol Chem 268 1908-1913 (1993).

Ogura, T., and Esumi, H. Nitric oxide synthase expression in human neuroblastoma cell line induced by cytokines. Mol Neurosci 7 853-856 (1996).

Michel, T., Xie, Q., and Nathan, C. Molecular biological analysis of nitric oxide synthases. 161-175 (1996).

Schmidt, H.H.H.W., Warner, T.D., Nakane, M., et al. Regulation and subcellular location of nitrogen oxide synthases in RAW264.7 macrophages. Mol Pharmacol 41 615-624 (1992).

Balligand, J., Ungureanu-Longrois, D., Simmons, W.W., et al. Cytokine-inducible nitric oxide synthase (iNOS) expression in cardiac myocytes. Characterization and regulation of iNOS expression and detection of iNOS activity in single cardiac cyocytes in vivo. J Biol Chem 269 27580-27588 (1994).

Walker, G., Pfeilschifter, J., and Kunz, D. Mechanism of suppression of inducible nitric-oxide synthase (iNOS) expression in interferon (IFN)-γ-stimulated RAW 264.7 cells by dexamethasone. Evidence for glucocorticoid-induced degradation of iNOS protein by calpain as a key step in post-transcriptional regulation. J Biol Chem 271 16679-16687 (1996).

Herencia, F., Ferrándiz, M.L., Ubeda, A., et al. Novel anti-inflammatory chalcone derivatives inhibit the induction of nitric oxide synthase and cyclooxygenase-2 in mouse peritoneal macrophages. FEBS Lett 453 129-134 (1999).

Hayes, J.D., and McLellan, L.I. Glutathione and glutathione-dependent enzymes represent a co-ordinately regulated defence against oxidative stress. Free Radic Res 31 273-300 (1999).

Smith, F.S., and Titheradge, M.A. Detection of NOS isoforms by western-blot analysis. Methods Mol Biol 100 171-180 (1998).

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