iNOS Antibody (4E5) - BSA Free

Western Blot: iNOS Antibody (4E5) [NBP2-22119] - Analysis using iNOS mouse mAb against Jurkat (1), Jurkat (2), A549 (3), HeLa (4), NIH3T3 (5) and MCF-7 (6) cell lysate.

Immunohistochemistry-Paraffin: iNOS Antibody (4E5) [NBP2-22119] - Analysis of paraffin-embedded breast cancer tissues using iNOS mouse mAb with DAB staining.

Flow Cytometry: iNOS Antibody (4E5) [NBP2-22119] - An intracellular stain was performed on A549 cells with NBP2-22119APC (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated with antibody at 1 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to allophycocyanin (APC).

Flow Cytometry: iNOS Antibody (4E5) [NBP2-22119] - An intracellular stain was performed on A549 cells with NBP2-22119PE (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated with antibody at 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to phycoerythrin (PE).

Western Blot: iNOS Antibody (4E5) [NBP2-22119] - Analysis using iNOS mAb against human iNOS (AA: 997-1058) recombinant protein. (Expected MW is 32.6 kDa)

Flow Cytometry: iNOS Antibody (4E5) - BSA Free [NBP2-22119] - An intracellular stain was performed on NIH3T3 cells with iNOS Antibody (4E5) NBP2-22119 (blue) and a matched isotype control MAB002 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (84540, Thermo Fisher).

Immunohistochemistry-Paraffin: iNOS Antibody (4E5) [NBP2-22119] - Analysis of paraffin-embedded liver cancer tissues using iNOS mouse mAb with DAB staining.

Flow Cytometry: iNOS Antibody (4E5) - BSA Free [NBP2-22119] - An intracellular stain was performed on Caco-2 cells with iNOS Antibody (4E5) NBP2-22119 (blue) and a matched isotype control MAB002 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (84540, Thermo Fisher).

ELISA: iNOS Antibody (4E5) [NBP2-22119] - Red: Control Antigen (100ng); Purple: Antigen (10ng); Green: Antigen (50ng); Blue: Antigen (100ng).

• Reactivity: Hu, Mu
• Applications: WB, ELISA, Flow, IHC
• Clone: 4E5
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

iNOS Antibody (4E5) - BSA Free Summary

• Immunogen: iNOS Antibody (4E5) was made to a purified recombinant fragment of human iNOS (C-terminus) expressed in E. coli. [UniProt# P35228]
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: NOS2
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • ELISA 1:10000
  • Flow (Intracellular)
  • Flow Cytometry 1:200-1:400
  • Immunohistochemistry reported in scientific literature (PMID 29891729)
  • Immunohistochemistry-Frozen reported in scientific literature (PMID 32780728)
  • Immunohistochemistry-Paraffin 1:200-1:1000
  • Western Blot 1:500-1:2000
• Application Notes: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Theoretical MW: 131 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein A purified

Alternate Names for iNOS Antibody (4E5) - BSA Free

• EC 1.14.13.39
• Hepatocyte NOS
• HEP-NOS
• Inducible NO synthase
• Inducible NOS
• iNOS
• nitric oxide synthase 2, inducible
• nitric oxide synthase 2A (inducible, hepatocytes)
• nitric oxide synthase, inducible
• nitric oxide synthase, macrophage
• NOS type II
• NOS
• NOS2
• NOS2A
• Peptidyl-cysteine S-nitrosylase NOS2

Background

Nitric oxide (NO) is a colorless, free radical gas 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 including the two constitutive isoforms: brain, bNOS, or neuronal NOS, nNOS (type I) and endothelial cell NOS, eNOS (type III); along with the inducible isoform, iNOS (type II). NOS catalyzes the oxidization of L-arginine to produce L-citrulline and NO, requiring the cofactors calmodulin, nicotinamide adenine dinucleotide phosphate (NADPH), flavin adenine dinucleotide (FAD), and flavin mononucleotide (FMN), heme, and tetrahydrobiopterin (1).

The 131 kDa enzyme, iNOS, is found in a variety of cell types including macrophages, hepatocytes, synoviocytes, and smooth muscle cells. While constitutively expressed in kidneys, in other tissues iNOS is induced by bacterial lipopolysaccharides (LPS), growth factors, and cytokines such as IFN-gamma, TNF, IL-1 and IL-2. iNOS is not regulated by the level of intracellular Ca2+ and is constantly active as a dimer when expressed. iNOS activity is elevated in a variety of diseases including atherosclerosis, heart failure, sepsis, solid tumors, and type 2 diabetes. Acting as a critical mediator of inflammation and apoptosis, iNOS inhibitors have been shown to alleviate obesity and stress inducted insulin resistance in mouse models (2,3).

References

1. Forstermann U, and Sessa WC. (2012) Nitric oxide synthases: regulation and function. Eur Heart J. 33(7): 829-837. PMID: 21890489

2. Aktan F. (2004) iNOS-mediated nitric oxide production and its regulation. Life Sci. 75(6):639-53. PMID: 15172174

3. Cinelli MA, Do HT, Miley GP, Silverman RB. (2020) Inducible nitric oxide synthase: Regulation, structure, and inhibition. Med Res Rev. 40(1):158-189. PMID: 31192483

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for iNOS Antibody (NBP2-22119)(40)

Publications using NBP2-22119

• Kong Y, Shi W, Zheng L et al. In situ delivery of a curcumin-loaded dynamic hydrogel for the treatment of chronic peripheral neuropathy Journal of controlled release : official journal of the Controlled Release Society 2023-04-08 [PMID: 37028453] (WB, Rat)
• Fan P, Yang Y, Liu T Et Al. Anti-atopic effect of Viola yedoensis ethanol extract against 2,4-dinitrochlorobenzene-induced atopic dermatitis-like skin dysfunction Journal of ethnopharmacology 2021-07-28 [PMID: 34332065]
• Cui J, Shan K, Yang Q et al. Prostaglandin E3 attenuates macrophage-associated inflammation and prostate tumour growth by modulating polarization Journal of cellular and molecular medicine 2021-05-13 [PMID: 33982835] (FLOW, Human)
• Selvaraj UM, Ujas TA, Kong X et al. Delayed diapedesis of CD8 T cells contributes to long-term pathology after ischemic stroke in male mice Brain, behavior, and immunity 2021-05-05 [PMID: 33964435] (IF/IHC, Mouse)
• Zhang J, Thakuri BKC, Zhao J et al. Long Noncoding RNA RUNXOR Promotes Myeloid-Derived Suppressor Cell Expansion and Functions via Enhancing Immunosuppressive Molecule Expressions during Latent HIV Infection Journal of immunology (Baltimore, Md. : 1950) 2021-05-01 [PMID: 33820854]
• Hu G, Su Y, Kang BH et al. High-throughput phenotypic screen and transcriptional analysis identify new compounds and targets for macrophage reprogramming Nature communications 2021-02-03 [PMID: 33536439] (FLOW, Mouse)
• Tyagi A, Sharma S, Wu K, et al. Nicotine promotes breast cancer metastasis by stimulating N2 neutrophils and generating pre-metastatic niche in lung Nature communications 2021-01-20 [PMID: 33473115] (FLOW, Human)
• Su S, Qin S, Xian X et al Interleukin-22 regulating Kupffer cell polarization through STAT3/Erk/Akt crosstalk pathways to extenuate liver fibrosis Life Sci 2020-11-01 [PMID: 33129875] (FLOW, Mouse)
  
  Details:
  Citation using the Alexa Fluor 532 version of this antibody.
• Feng J, Chen S, Li S et al The association between monocytic myeloid-derived suppressor cells levels and the anti-tumor efficacy of anti-PD-1 therapy in NSCLC patients Transl Oncol 2020-09-13 [PMID: 32920330]
  
  Details:
  Citation using the PE version of this antibody.
• Chen L, Wang D, Liu X Et Al. Micro-galvanic effects of silver-containing titanium implants regulate the immune responses via activating voltage-gated calcium channels in macrophages Chemical Engineering Journal 2021-01-01 (ICC/IF)
• Wilkinson HN, Reubinoff B, Shveiky D et al. Epithelial arginase-1 is a key mediator of age-associated delayed healing in vaginal injury Frontiers in endocrinology 2022-08-11 [PMID: 36034415] (IHC-P, Rat)
• Shueng Pw, Yu Ly, Chiu Hc Et Al. Early phago-/endosomal escape of platinum drugs via ROS-responsive micelles for dual cancer chemo/immunotherapy Biomaterials 2021-07-07 [PMID: 34252800] (IHC-P)
• Goldfarb S, Fainstein N, Ben-Hur T Electroconvulsive stimulation attenuates chronic neuroinflammation JCI Insight 2020-09-03 [PMID: 32780728] (IHC-Fr, Mouse)
• Kim G, Hyun C The hyaluronan synthesis inhibitor 7-hydroxy-4-methylcoumarin inhibits LPS-induced inflammatory response in RAW 264.7 macrophage cells Journal of Applied Biological Chemistry 2021-09-30
• Zhang W, Chen J, Guo W et al. WKYMVm/FPR2 Alleviates Spinal Cord Injury by Attenuating the Inflammatory Response of Microglia Mediators of Inflammation 2022-07-27 [PMID: 35935810] (IHC-P, Rat)
• Chen Y, Li P, Zhen R et al. Effects of niacin on intestinal epithelial Barrier, intestinal Immunity, and microbial community in weaned piglets challenged by PDCoV International immunopharmacology 2022-08-01 [PMID: 35921778] (WB, Mouse)
• Yan B, Tan J, Zhang H et al. Constructing fluorine-doped Zr-MOF films on titanium for antibacteria, anti-inflammation, and osteogenesis Materials Science and Engineering: C 2022-02-01 [PMID: 35581071]
• Ganz T, Fainstein N, Elad A et al. Microbial pathogens induce neurodegeneration in Alzheimer's disease mice: protection by microglial regulation Journal of neuroinflammation 2022-01-06 [PMID: 34991645] (IHC-Fr, Mouse)
• MartIn-Cordero L, Galvez I, Hinchado MD, Ortega E Influence of Obesity and Exercise on beta 2-Adrenergic-Mediated Anti-Inflammatory Effects in Peritoneal Murine Macrophages Biomedicines 2020-11-30 [PMID: 33266248] (FLOW, Human)
• Chu C, Hu C, Rung S et al. ARG1 could be expressed by human, but not represent for repair bioRxiv 2020-07-03 (FLOW, Human)
• Beielstein AC, Izquierdo E, Blakemore S, Nickel N et Al. Macrophages are activated toward phagocytic lymphoma cell clearance by pentose phosphate pathway inhibition Cell Rep Med 2024-11-27 [PMID: 39603243]
• Jim Middelburg, Soroush Ghaffari, Tom A W Schoufour, Marjolein Sluijter, Gaby Schaap, Büsra Göynük, Benedetta M Sala, Lejla Al-Tamimi, Ferenc Scheeren, Kees L M C Franken, Jimmy J L L Akkermans, Birol Cabukusta, Simone A Joosten, Ian Derksen, Jacques Neefjes, Sjoerd H van der Burg, Adnane Achour, Ruud H M Wijdeven, Jon Weidanz, Thorbald van Hall The MHC-E peptide ligands for checkpoint CD94/NKG2A are governed by inflammatory signals, whereas LILRB1/2 receptors are peptide indifferent. Cell reports 2024-01-01 [PMID: 38048225]
• Zhang Y, Li W, Guo S et al. FBXO22 Mediates the NGF/TRKA Signaling Pathway in Bone Metastases in Prostate Cancer The American journal of pathology 2023-06-08 [PMID: 37301536] (IHC, Mouse)
• You M, Song Y, Chen J et al. Combined exposure to benzo(a)pyrene and dibutyl phthalate aggravates pro-inflammatory macrophage polarization in spleen via pyroptosis involving cathepsin B The Science of the total environment 2023-04-14 [PMID: 37061049]
• Zhao S, Zhou L, Wang Q et al. Elevated branched-chain amino acid promotes atherosclerosis progression by enhancing mitochondrial-to-nuclear H2O2-disulfide HMGB1 in macrophages Redox biology 2023-04-05 [PMID: 37058999] (IHC-P, Mouse)
• Cheng H, Sung W, Tseng C, Lee L Bacillus amyloliquefaciens exopolysaccharides promote the defensive and protective functions of intestinal epithelial cells AUTHOREA 2023-01-01
• He Z, Greven J, Shi Y et al. Extracellular vesicles derived from endothelial cells modulate macrophage phenotype in vitro European journal of medical research 2023-11-09 [PMID: 37946271]
• Zheng X, Chen L, Tan J et al. Effect of micro/nano-sheet array structures on the osteo-immunomodulation of macrophages Regenerative biomaterials 2022-10-04 [PMID: 36284748]
• Roy R, Zayas J, Mohamed MF et al. IL-10 Dysregulation Underlies Chemokine Insufficiency, Delayed Macrophage Response, and Impaired Healing in Diabetic Wounds Journal of Investigative Dermatology 2022-03-01 [PMID: 34517005] (Immunohistochemistry-Paraffin, Human)
• Marques RM, Gonzalez-Nunez M, Walker ME et al. Loss of 15-lipoxygenase disrupts T(reg) differentiation altering their pro-resolving functions Cell Death & Differentiation 2021-11-01 [PMID: 34040168] (In vivo assay)
• Hu C, Chu C, Liu L et al. Dissecting the microenvironment around biosynthetic scaffolds in murine skin wound healing Science Advances 2021-05-28 [PMID: 34039601] (Flow Cytometry, In vivo assay)
• Hor‐Yue Tan, Ning Wang, Cheng Zhang, Yau‐Tuen Chan, Man‐Fung Yuen, Yibin Feng Lysyl Oxidase‐Like 4 Fosters an Immunosuppressive Microenvironment During Hepatocarcinogenesis Hepatology (Baltimore, Md.) 2021-05-21 [PMID: 33068461]
• Nishri Y, Hampton D, Ben-Shushan E et al. Continuous Immune-Modulatory Effects of Human Olig2+ Precursor Cells Attenuating a Chronic-Active Model of Multiple Sclerosis Mol. Neurobiol. 2019-10-28 [PMID: 31656989] (Mouse)
• Galvez I, Martin-Cordero L, Hinchado MD Anti-inflammatory effect of beta-2 adrenergic stimulation on circulating monocytes with a pro-inflammatory state in high-fat diet-induced obesity Brain Behav. Immun. 2019-05-03 [PMID: 31055173] (FLOW, Mouse)
• Bastian C, Zaleski J, Stahon K et al. NOS3 Inhibition Confers Post-Ischemic Protection to Young and Aging White Matter Integrity by Conserving Mitochondrial Dynamics and Miro-2 Levels. J. Neurosci. 2018-07-11 [PMID: 29891729] (IF/IHC, Mouse)
• Pistorius K, Souza PR, De Matteis R et al. PDn-3 DPA Pathway Regulates Human Monocyte Differentiation and Macrophage Function Cell Chem Biol May 16 2018 [PMID: 29805036] (Flow) Cell Chem Biol. 2018-05-24 [PMID: 29805036] (FLOW)
  
  Details:
  Citation using the Alexa Fluor 647 form of this antibody.
• Omlor AJ, Le DD, Schlicker J et al. Local Effects on Airway Inflammation and Systemic Uptake of 5 nm PEGylated and Citrated Gold Nanoparticles in Asthmatic Mice. Small. 2016-12-23 [PMID: 28009478]
• Okuyama N, Matsuda S, Yamashita A et al. Human Coronary Thrombus Formation Is Associated With Degree of Plaque Disruption and Expression of Tissue Factor and Hexokinase II. Circ. J. 2015-09-03 [PMID: 26346032] (IHC-P, Human)
• Padgett Le, Burg Ar, Lei W, Tse Hm. Loss of nADPH Oxidase-Derived Superoxide Skews Macrophage Phenotypes to Delay Type 1 Diabetes. Diabetes. 2014-10-06 [PMID: 25288672] (WB)
• Wang Y, Wang W, Zhang Y et al. Clinical significance of a myeloperoxidase gene polymorphism and inducible nitric oxide synthase expression in cirrhotic patients with hepatopulmonary syndrome. J Huazhong Univ Sci Technolog Med Sci. 2010-08-01 [PMID: 20714866] (IHC-P, Human)

Review for iNOS Antibody (NBP2-22119) (1)

Average Rating: 3

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer WB Human 01/01/2017

Summary

• Application: Western Blot
• Sample Tested: Placental tissue
• Species: Human
• Lot: A-4

Comments

• Comments: Non induced, placental tissue on delivery.

FAQs for iNOS Antibody (NBP2-22119). (Showing 1 - 2 of 2 FAQs).

1. We need to evaluate the level of iNOS in Rat sciatic nerve by Western Blot. I see that you have few antibodies suitable for that, could you please recommend the best what you have for our purposes?
   • We have 4 iNOS antibodies that are suitable for your experiment. The best choice for you will depend on your specific preferences, in this case the main difference will be the immunogen location. If the immunogen is not important to you, then I would recommend choosing NB300-605 as it gives you the best value and we are able to provide the exact immunogen.
2. We are having trouble figuring out the correct dilution for IHC. What are the recommended dilutions for this product’s applications?
   • The dilutions for IHC can vary by catalog number with suggested ranges of anywhere from 1:10 to 1:1000 across our iNOS antibody products. Our recommended dilutions for each application are listed in our products’ data sheets, which can be found on their product webpages. If no dilution guidelines are listed, we recommend running preliminary tests to find the optimal dilution for your experiments.

Bioinformatics

• Gene Symbol: NOS2
• Entrez:
  • 4843(Human)
• Uniprot:
  • P35228()