iNOS Monoclonal Antibody (CXNFT), Brilliant Ultra Violet™ 737, eBioscience™, Invitrogen™

Description

• This CXNFT monoclonal antibody reacts to mouse NOS2 (inducible NOS, iNOS)
• Nitric oxide synthase enzymes catalyze the formation of nitric oxide from L-arginine through an NADPH- and oxygen-dependent mechanism
• There are three isoforms of NOS that are encoded by three separate genes
• NOS1 and NOS3are constitutively expressed, while NOS2 is induced in response to bacterial endotoxins and inflammatory cytokines such as IFN gamma and TNF alpha
• NOS2 is expressed by myeloid-derived suppressor cells and M1 macrophages but not alternatively activated M2 macrophages
• NOS enzymes are functionally active only when they form homodimers, and dimerization of NOS2 occurs at steady-state concentrations of free Ca2+ such that NOS2 is functionally active when it is produced
• Applications Reported: This CXNFT antibody has been reported for use in intracellular staining followed by flow cytometric analysis
• Applications Tested: This CXNFT antibody has been tested by intracellular staining followed by flow cytometric analysis of stimulated mouse thioglycolate-elicited peritoneal macrophages using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00)
• This may be used at less than or equal to 0.125 μg per test
• Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test
• Brilliant Ultraviolet 737 is a tandem dye that emits at 732 nm and is intended for use on cytometers equipped with an ultraviolet (355 nm) laser
• iNOS (Inducible Nitric oxide, NO, NOS) is an inorganic, gaseous free radical that carries a variety of messages between cells
• Vasorelaxation, neurotransmission and cytotoxicity can all be potentiated through cellular response to NO
• NO production is mediated by members of the nitric oxide synthase (NOS) family
• iNOS is expressed in liver and inducible by a combination of lipopolysaccharide and certain cytokines
• NOS catalyzes the oxidization of L-arginine to produce L-citrulline and NO
• Two constitutive isoforms, brain or neuronal NOS (b or nNOS, type I) and endothelial cell NOS (eNOS, type III), and one inducible isoform (iNOS, type II), have been cloned
• All NOS isoforms contain calmodulin, nicotinamide adenine dinucleotide phosphate (NADPH), flavin adenine dinucleotide (FAD), and flavin mononucleotide (FMN) binding domains
• iNOS is found in a variety of cell types including macrophages, hepatocytes, synoviocytes, and smooth muscle cells
• Cytokines such as interferon-gamma (IFN), tumor necrosis factor (TNF), interleukin-1 and -2, and lipopolysaccarides (LPS) cause an increase in iNOS mRNA, protein, and activity levels
• Protein kinase C-stimulating agents exhibit the same effect on iNOS activity
• After cytokine induction, iNOS exhibits a delayed activity response which is then followed by a significant increase in NO production over a long period of time
• Three related iNOS pseudogenes are located within the Smith-Magenis syndrome region on chromosome 17
• Diseases associated with iNOS dysfunction include achalasia and impotence.