Invitrogen

iNOS Monoclonal Antibody (CXNFT), Alexa Fluor™ 488, eBioscience™

Advanced Verification

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated.

14 References

View all (65) iNOS antibodies

Datasheet

Protocols

Questions & Answers

Datasheet

Protocols

Questions & Answers

Antibody Testing Data (3)
Advanced Verification (2)

iNOS Antibody in Immunocytochemistry (ICC/IF)

FIGURE: 1 / 5

iNOS Antibody (53-5920-82) in ICC/IF

Immunofluorescence analysis of Nitric oxide synthase, inducible, was performed using 70% confluent log phase RAW 264.7 treated with LPS (1 µg/mL for 16h). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with iNOS Monoclonal Antibody (CXNFT), Alexa Fluor 488, eBioscience™ (Product # 53-5920-82) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight (Panel a: Green). Nuclei (Pa... View More

Product Image Gallery

iNOS Antibody — Detection of altered expression of the target protein by cell treatment demonstrates antibody specificity. Immunofluorescence analysis using iNOS Monoclonal Antibody (CXNFT), Alexa Fluor 488, eBioscience™ (Product # 53-5920-82), shows increased expression of iNOS upon treatment of RAW264.7 cells with LPS. {TM}

Product Details

53-5920-82

Applications

Tested Dilution

Publications

Western Blot (WB)

1:1,000

Immunocytochemistry (ICC/IF)

1:100

View 1 publication 1 publication

Flow Cytometry (Flow)

0.06 µg/test

View 13 publications 13 publications

Product Specifications

Species Reactivity

Mouse

Published species

Human, Mouse

Host/Isotype

Rat / IgG2a, kappa

Recommended Isotype Control

Rat IgG2a kappa Isotype Control (eBR2a), Alexa Fluor™ 488, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

CXNFT

Conjugate

Alexa Fluor™ 488 Alexa Fluor™ 488 Alexa Fluor™ 488

Excitation/Emission Max

499/520 nm View spectra

Form

Liquid

Concentration

0.5 mg/mL

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

AB_2574423

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 stimulated mouse thioglycolate-elicited peritoneal exudate cells using the intracellular Fixation and Permeabilization Buffer Set (Product # 88-8824-00) and protocol. 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.

Excitation: 488 nm; Emission: 519 nm; Laser: Blue 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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Product References

Immune Deviation in the Decidua During Term and Preterm Labor.

Frontiers in immunology

Zha Y,Liu H,Lin X,Yu L,Gao P,Li Y,Wu M,Gong X,Bian X,Kang Q,Zhi P,Dang X,Wang J,Feng L,Qiao F,Huang Y,Zeng W

53-5920-82 was used in flow cytometry to demonstrate significantly decreased frequencies of M2 cells and elevated M1/M2 ratio in the PIL group compared to that in the PNL group in both decidua basalis and parietalis, whereas no significant differences were found between the above two groups in both sites in terms of the polarization status of Th cells.

Tue Jun 28 00:00:00 EDT 2022

Cold exposure protects from neuroinflammation through immunologic reprogramming.

Cell metabolism

Spiljar M,Steinbach K,Rigo D,Suárez-Zamorano N,Wagner I,Hadadi N,Vincenti I,Page N,Klimek B,Rochat MA,Kreutzfeldt M,Chevalier C,Stojanović O,Bejuy O,Colin D,Mack M,Cansever D,Greter M,Merkler D,Trajkovski M

53-5920-82 was used in Flow Cytometry, Immunocytochemistry, Western Blot to propose a mechanistic link between environmental temperature and neuroinflammation and suggest competition between cold-induced metabolic adaptations and autoimmunity as energetic trade-off beneficial for the immune-mediated diseases.

Tue Nov 02 00:00:00 EDT 2021

miR-155-5p regulates macrophage M1 polarization and apoptosis in the synovial fluid of patients with knee osteoarthritis.

Experimental and therapeutic medicine

Li GS,Cui L,Wang GD

53-5920 was used in Flow cytometry/Cell sorting to suggest that miR-155-5p may serve as a potential therapeutic target for KOA.

Fri Jan 01 00:00:00 EST 2021

Redox lipid reprogramming commands susceptibility of macrophages and microglia to ferroptotic death.

Nature chemical biology

Kapralov AA,Yang Q,Dar HH,Tyurina YY,Anthonymuthu TS,Kim R,St Croix CM,Mikulska-Ruminska K,Liu B,Shrivastava IH,Tyurin VA,Ting HC,Wu YL,Gao Y,Shurin GV,Artyukhova MA,Ponomareva LA,Timashev PS,Domingues RM,Stoyanovsky DA,Greenberger JS,Mallampalli RK,Bahar I,Gabrilovich DI,Bayır H,Kagan VE

53-5920 was used in Immunocytochemistry to discover that inducible nitric oxide synthase (iNOS)/NO-enrichment of activated M1 (but not alternatively activated M2) macrophages/microglia modulates susceptibility to ferroptosis.

Sun Mar 01 00:00:00 EST 2020

Macrophage polarization plays roles in bone formation instructed by calcium phosphate ceramics.

Journal of materials chemistry. B

Li M,Guo X,Qi W,Wu Z,de Bruijn JD,Xiao Y,Bao C,Yuan H

53-5920 was used in Flow cytometry/Cell sorting to investigate the roles of macrophages in material-instructed bone formation.

Wed Mar 04 00:00:00 EST 2020

Enhanced alleviation of aGVHD by TGF-β1-modified mesenchymal stem cells in mice through shifting MΦ into M2 phenotype and promoting the differentiation of Treg cells.

Journal of cellular and molecular medicine

Wu R,Liu C,Deng X,Chen L,Hao S,Ma L

53-5920 was used in Flow cytometry/Cell sorting to study the mechanism by which mesenchymal stem cells can treat steroid resistant acute graft-versus-host disease.

Wed Jan 01 00:00:00 EST 2020

Mannose receptor high, M2 dermal macrophages mediate nonhealing Leishmania major infection in a Th1 immune environment.

The Journal of experimental medicine

Lee SH,Charmoy M,Romano A,Paun A,Chaves MM,Cope FO,Ralph DA,Sacks DL

53-5920 was used in Flow cytometry/Cell sorting to study the origin and functional specialisation of dermal macrophages in cutaneous infections.

Tue Jan 02 00:00:00 EST 2018

Eosinophil recruitment is dynamically regulated by interplay among lung dendritic cell subsets after allergen challenge.

Nature communications

Yi S,Zhai J,Niu R,Zhu G,Wang M,Liu J,Huang H,Wang Y,Jing X,Kang L,Song W,Shi Y,Tang H

53-5920 was used in Flow cytometry/Cell sorting to suggest that different lung antigen-presenting cells modulate lung cDC1-mediated eosinophil recruitment dynamically.

Mon Sep 24 00:00:00 EDT 2018

Rediscovering peritoneal macrophages in a murine endometriosis model.

Human reproduction (Oxford, England)

Yuan M,Li D,An M,Li Q,Zhang L,Wang G

53-5920 was used in Flow Cytometry to study the role of large and small peritoneal macrophages in endometriosis.

Sun Jan 01 00:00:00 EST 2017

Regulatory T Cells Promote Myositis and Muscle Damage in Toxoplasma gondii Infection.

Journal of immunology (Baltimore, Md. : 1950)

Jin RM,Blair SJ,Warunek J,Heffner RR,Blader IJ,Wohlfert EA

53-5920 was used in Flow cytometry/Cell sorting to study how the tissue environment established by Toxoplasma gondii leads to a pathogenic role for regulatory T cells.

Sun Jan 01 00:00:00 EST 2017

Activation of the NLRP3 Inflammasome Is Associated with Valosin-Containing Protein Myopathy.

Inflammation

Nalbandian A,Khan AA,Srivastava R,Llewellyn KJ,Tan B,Shukr N,Fazli Y,Kimonis VE,BenMohamed L

53-5920 was used in Flow cytometry/Cell sorting to investigate whether the NLRP3 inflammasome plays a role in valosin-containing protein-associated diseases, which classically affects the muscles, bones, and brain.

Wed Feb 01 00:00:00 EST 2017

Ginsenoside Rd alleviates mouse acute renal ischemia/reperfusion injury by modulating macrophage phenotype.

Journal of ginseng research

Ren K,Jin C,Ma P,Ren Q,Jia Z,Zhu D

53-5920 was used in Flow cytometry/Cell sorting to investigate the effect of GSRd on renal ischemia/reperfusion injury (IRI) and macrophage functional status, showing that it alleviates injury by modulating macrophage phenotype.

Fri Apr 01 00:00:00 EDT 2016

Commensal Gram-positive bacteria initiates colitis by inducing monocyte/macrophage mobilization.

Mucosal immunology

Nakanishi Y,Sato T,Ohteki T

Thu Jan 01 00:00:00 EST 2015

Advanced Glycation End Products Enhance Macrophages Polarization into M1 Phenotype through Activating RAGE/NF-κB Pathway.

BioMed research international

Jin X,Yao T,Zhou Z,Zhu J,Zhang S,Hu W,Shen C

53-5920 was used in Flow cytometry/Cell sorting to assess the effects of AGEs on macrophage polarization.

Fri Mar 18 00:00:00 EDT 2016

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

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Gene Aliases: i-NOS; iNOS; Inosl; Nos-2; NOS-II; Nos2; Nos2a

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UniProt ID: (Mouse) P29477

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Entrez Gene ID: (Mouse) 18126

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Function(s)

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

Process(es)

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

Performance Guarantee

If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*

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iNOS Monoclonal Antibody (CXNFT), Alexa Fluor™ 488, eBioscience™

iNOS Antibody (53-5920-82) in ICC/IF

Product Details

53-5920-82

Western Blot (WB)

Immunocytochemistry (ICC/IF)

Flow Cytometry (Flow)

Species Reactivity

Published species

Host/Isotype

Recommended Isotype Control

Class

Type

Clone

Conjugate

Excitation/Emission Max

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

Shipping conditions

RRID

Product Specific Information

Target Information

How to use the Panel Builder

Bioinformatics

Performance Guarantee

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