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iNOS Monoclonal Antibody (CXNFT), PE-eFluor™ 610, eBioscience™
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FIGURE: 1 / 30
iNOS Antibody (61-5920-82) in Flow
Mouse thioglycolate-elicited peritoneal exudate cells were stimulated overnight with LPS then surface stained with Anti-Mouse CD11b eFluor® 450 (Product # 48-0112-82) followed by fixation and permeabilization with the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol. The cells were then intracellularly stained with 0.03 µg of Rat IgG2a K Isotype Control PE-eFluor® 610 (Product # 61-4321-82) (left) or 0.03 µg of Anti-Mouse NOS2 PE-eFluor® 610 (right). Total viable cells, as determined by Fixable Viabi... View More
Product Details
61-5920-82
Applications
Tested Dilution
Publications
Product Specifications
Species Reactivity
Mouse
Published species
Mouse
Host/Isotype
Rat
/ IgG2a, kappa
Recommended Isotype Control
Class
Monoclonal
Type
Antibody
Clone
CXNFT
Conjugate
Excitation/Emission Max
565/606 nm
View spectra
Form
Liquid
Concentration
0.2 mg/mL
Amount
100 µg
Purification
Affinity chromatography
Storage buffer
PBS, pH 7.2
Contains
0.09% sodium azide
Storage conditions
4°C, store in dark, DO NOT FREEZE!
Shipping conditions
Ambient (domestic); Wet ice (international)
RRID
Product Specific Information
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 (neuronal NOS, nNOS) and NOS3 (endothelial NOS, eNOS) are 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 and flow cytometric analysis of LPS-stimulated mouse thioglycolate-elicited peritoneal exudate cells using the Intracellular Fixation and Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Please refer to BestProtocols®: Protocol A: Two step protocol for (cytoplasmic) intracellular proteins located under the Resources Tab online. The Foxp3/Transcription Factor Staining Buffer Set (Product # 00-5523-00) may also be used with similar results. This can be used at less than or equal to 0.06 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
PE-eFluor® 610 can be excited with laser lines from 488-561 nm and emits at 607 nm. We recommend using a 610/20 band pass filter (equivalent to PE-Texas Red®). Please make sure that your instrument is capable of detecting this fluorochome.
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 488-561 nm; Emission: 607 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.
Filtration: 0.2 µm post-manufacturing filtered.
Target Information
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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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Bioinformatics
Protein Aliases: hepatocytes; inducible nitric oxide synthase; Inducible NO synthase; Inducible NOS; MAC-NOS; Macrophage NOS; nitric oxide synthase 2, inducible, macrophage; Nitric oxide synthase, inducible; nitric oxide synthase-inducible; NOS type II; OTTMUSP00000000202; Peptidyl-cysteine S-nitrosylase NOS2
Gene Aliases: i-NOS; iNOS; Inosl; Nos-2; NOS-II; Nos2; Nos2a
UniProt ID: (Mouse) P29477
Entrez Gene ID: (Mouse) 18126
actin binding
NADPH-hemoprotein reductase activity
nitric-oxide synthase activity
receptor binding
iron ion binding
protein binding
calmodulin binding
beta-catenin binding
cAMP-dependent protein kinase regulator activity
FMN binding
oxidoreductase activity
protein kinase binding
heme binding
tetrahydrobiopterin binding
arginine binding
protein homodimerization activity
cadherin binding
metal ion binding
flavin adenine dinucleotide binding
nitric-oxide synthase binding
Hsp90 protein binding
oxidoreductase
ovulation from ovarian follicle
response to hypoxia
arginine catabolic process
superoxide metabolic process
nitric oxide biosynthetic process
inflammatory response
nitric oxide mediated signal transduction
negative regulation of gene expression
peptidyl-cysteine S-nitrosylation
positive regulation of guanylate cyclase activity
response to lipopolysaccharide
cellular response to drug
regulation of cell proliferation
negative regulation of protein catabolic process
defense response to bacterium
negative regulation of blood pressure
regulation of protein kinase activity
positive regulation of vasodilation
regulation of insulin secretion
positive regulation of killing of cells of other organism
oxidation-reduction process
cellular response to lipopolysaccharide
cellular response to interferon-gamma
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