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Technical Data Sheet

NM5751

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iNOS (C-terminal region) M575
Mouse Monoclonal IgG2a
Price
Size
Species Reactivity
MW

$295
100 μl
Hu, Rt, Ms
130 kDa

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Enlarge

Western blot analysis of mouse macrophage (RAW264.7) untreated (lane 1) or treated with IFNγ (10 ng/ml) and LPS (1µg/ml) for 12 hrs (lanes 2). The blot was probed with mouse monoclonal anti-iNOS M575 at 1:1000 (lanes 1 & 2).

Application
Dilution


ELISA
1:2000


ICC
1:200


IP
1:100


WB
1:1000



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
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Background
Nitric oxide (NO) has a broad range of biological activities and is implicated in signaling pathways in phylogenetically diverse species. Nitric oxide synthases (NOS), the enzymes responsible for synthesis of NO, are homodimers whose monomers are themselves two fused enzymes: a cytochrome reductase and a cytochrome that requires three cosubstrates (L-arginine, NADPH, and oxygen) and five cofactors or prosthetic groups (FAD, FMN, calmodulin, tetrahydrobiopterin, and heme). Several distinct NOS isoforms are produced from three distinct genes. These include two constitutive Ca2+/CaM-dependent forms of NOS: nNOS (also designated bNOS, NOS-I), whose activity was first identified in neurons, and eNOS (also designated ecNOS, NOS-III) first identified in endothelial cells. The inducible form of NOS, iNOS (also designated NOS-II), is Ca2+ independent and is expressed in a broad range of cell types. This form of NOS is induced after stimulation with cytokines and exposure to microbial products.


Background References
Xie, Q.W. et al. (1992) Science 256:225.
Kleinert, H. et al. (2003) Biol Chem. 384(10-11):1343.
Musicki, B. et al. (2005) Proc. Natl. Acad. Sci.102(33):11870.
Immunogen
Clone (M575) was generated from a recombinant protein that included amino acid residues within the C-terminal region of human iNOS. The human iNOS sequence used has high homology with similar regions in rat and mouse iNOS.
Buffer and Storage
Mouse monoclonal antibody purified with protein A chromatography is supplied in 100µl phosphate-buffered saline, 50% glycerol,
1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
Specificity
The antibody detects a 130 kDa* protein on SDS-PAGE immunoblots of mouse macrophages (RAW264.7) treated with IFNγ and LPS, or J774A.1 cells treated with LPS only.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.
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