Product: iNOS Antibody
Catalog: AF0199
Source: Rabbit
Application: WB, IHC, IF/ICC, ELISA(peptide)
Reactivity: Human, Mouse, Rat
Mol.Wt.: 130kD; 131kD(Calculated).
Uniprot: P35228
RRID: AB_2833391

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Product Info

Source:
Rabbit
Application:
WB: 1:500-1:3000, IHC: 1:50-1:200, IF/ICC: 1:100-1:500, ELISA(peptide) 1:20000-1:40000
*The optimal dilutions should be determined by the end user.
Reactivity:
Human,Mouse,Rat
Clonality:
Polyclonal
Specificity:
iNOS Antibody detects endogenous levels of total iNOS.
RRID:
AB_2833391
Cite Format: Affinity Biosciences Cat# AF0199, RRID:AB_2833391.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

HEP-NOS; Hepatocyte NOS; HEPNOS; inducible; Inducible nitric oxide synthase; Inducible NO synthase; Inducible NOS; iNOS; MAC NOS; Macrophage NOS; Nitric oxide synthase 2 inducible; Nitric oxide synthase 2 inducible macrophage; nitric oxide synthase 2A (inducible, hepatocytes); Nitric oxide synthase; Nitric oxide synthase inducible; nitric oxide synthase, macrophage; NOS 2; NOS; Nos II; NOS type II; nos2; NOS2_HUMAN; NOS2A; NOS2A, Inducible, Hepatocyte; Peptidyl-cysteine S-nitrosylase NOS2;

Immunogens

Immunogen:

A synthesized peptide derived from human iNOS, corresponding to a region within C-terminal amino acids.

Uniprot:
Gene(ID):
Expression:
P35228 NOS2_HUMAN:

Expressed in the liver, retina, bone cells and airway epithelial cells of the lung. Not expressed in the platelets.

Description:
iNOS Produces nitric oxide (NO) which is a messenger molecule with diverse functions throughout the body. In macrophages, NO mediates tumoricidal and bactericidal actions. Belongs to the NOS family. Induced by endotoxins and cytokines. Induced by interferon gamma acting synergistically with lipopolysaccharide, tumor necrosis factor alpha or interleukin-1 beta. Homodimer. Binds SLC9A3R1. 2 isoforms of the human protein are produced by alternative splicing.
Sequence:
MACPWKFLFKTKFHQYAMNGEKDINNNVEKAPCATSSPVTQDDLQYHNLSKQQNESPQPLVETGKKSPESLVKLDATPLSSPRHVRIKNWGSGMTFQDTLHHKAKGILTCRSKSCLGSIMTPKSLTRGPRDKPTPPDELLPQAIEFVNQYYGSFKEAKIEEHLARVEAVTKEIETTGTYQLTGDELIFATKQAWRNAPRCIGRIQWSNLQVFDARSCSTAREMFEHICRHVRYSTNNGNIRSAITVFPQRSDGKHDFRVWNAQLIRYAGYQMPDGSIRGDPANVEFTQLCIDLGWKPKYGRFDVVPLVLQANGRDPELFEIPPDLVLEVAMEHPKYEWFRELELKWYALPAVANMLLEVGGLEFPGCPFNGWYMGTEIGVRDFCDVQRYNILEEVGRRMGLETHKLASLWKDQAVVEINIAVLHSFQKQNVTIMDHHSAAESFMKYMQNEYRSRGGCPADWIWLVPPMSGSITPVFHQEMLNYVLSPFYYYQVEAWKTHVWQDEKRRPKRREIPLKVLVKAVLFACMLMRKTMASRVRVTILFATETGKSEALAWDLGALFSCAFNPKVVCMDKYRLSCLEEERLLLVVTSTFGNGDCPGNGEKLKKSLFMLKELNNKFRYAVFGLGSSMYPRFCAFAHDIDQKLSHLGASQLTPMGEGDELSGQEDAFRSWAVQTFKAACETFDVRGKQHIQIPKLYTSNVTWDPHHYRLVQDSQPLDLSKALSSMHAKNVFTMRLKSRQNLQSPTSSRATILVELSCEDGQGLNYLPGEHLGVCPGNQPALVQGILERVVDGPTPHQTVRLEALDESGSYWVSDKRLPPCSLSQALTYFLDITTPPTQLLLQKLAQVATEEPERQRLEALCQPSEYSKWKFTNSPTFLEVLEEFPSLRVSAGFLLSQLPILKPRFYSISSSRDHTPTEIHLTVAVVTYHTRDGQGPLHHGVCSTWLNSLKPQDPVPCFVRNASGFHLPEDPSHPCILIGPGTGIAPFRSFWQQRLHDSQHKGVRGGRMTLVFGCRRPDEDHIYQEEMLEMAQKGVLHAVHTAYSRLPGKPKVYVQDILRQQLASEVLRVLHKEPGHLYVCGDVRMARDVAHTLKQLVAAKLKLNEEQVEDYFFQLKSQKRYHEDIFGAVFPYEAKKDRVAVQPSSLEMSAL

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
0
Horse
0
Bovine
0
Sheep
0
Dog
0
Xenopus
0
Zebrafish
0
Chicken
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P35228 As Substrate

Site PTM Type Enzyme
S50 Phosphorylation
S70 Phosphorylation
T77 Phosphorylation
S112 Phosphorylation
S124 Phosphorylation
T126 Phosphorylation
Y151 Phosphorylation P12931 (SRC)
Y267 Phosphorylation
Y270 Phosphorylation
S425 Phosphorylation
Y446 Phosphorylation
Y451 Phosphorylation
S453 Phosphorylation
K516 Ubiquitination
Y575 Phosphorylation
K604 Acetylation
R670 Methylation
S745 Phosphorylation P27361 (MAPK3)
Y868 Phosphorylation
S892 Phosphorylation
S898 Phosphorylation
Y1055 Phosphorylation P12931 (SRC)
Y1123 Phosphorylation
Y1134 Phosphorylation

Research Backgrounds

Function:

Produces nitric oxide (NO) which is a messenger molecule with diverse functions throughout the body. In macrophages, NO mediates tumoricidal and bactericidal actions. Also has nitrosylase activity and mediates cysteine S-nitrosylation of cytoplasmic target proteins such PTGS2/COX2 (By similarity). As component of the iNOS-S100A8/9 transnitrosylase complex involved in the selective inflammatory stimulus-dependent S-nitrosylation of GAPDH on 'Cys-247' implicated in regulation of the GAIT complex activity and probably multiple targets including ANXA5, EZR, MSN and VIM. Involved in inflammation, enhances the synthesis of proinflammatory mediators such as IL6 and IL8.

PTMs:

Polyubiquitinated; mediated by SPSB1, SPSB2 and SPSB4, leading to proteasomal degradation.

Subcellular Location:

Cytoplasm>Cytosol.
Note: Localizes as discrete foci scattered throughout the cytosol and in the presence of SPSB1 and SPSB4, exhibits a more diffuse cytosolic localization.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Expressed in the liver, retina, bone cells and airway epithelial cells of the lung. Not expressed in the platelets.

Subunit Structure:

Homodimer. Interacts with SLC9A3R1. Interacts with GAPDH; induced by oxidatively-modified low-densitity lipoprotein (LDL(ox)). Interacts with S100A8 and S100A9 to form the iNOS-S100A8/9 transnitrosylase complex. Interacts with SPSB1, SPSB2 and SPSB4. Interacts with ELOC and CUL5 in the presence of SPSB1 or SPSB2 or SPSB4.

Family&Domains:

Belongs to the NOS family.

Research Fields

· Cellular Processes > Transport and catabolism > Peroxisome.   (View pathway)

· Environmental Information Processing > Signal transduction > Calcium signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Apelin signaling pathway.   (View pathway)

· Human Diseases > Infectious diseases: Bacterial > Salmonella infection.

· Human Diseases > Infectious diseases: Bacterial > Pertussis.

· Human Diseases > Infectious diseases: Parasitic > Leishmaniasis.

· Human Diseases > Infectious diseases: Parasitic > Chagas disease (American trypanosomiasis).

· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.

· Human Diseases > Infectious diseases: Parasitic > Amoebiasis.

· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Small cell lung cancer.   (View pathway)

· Metabolism > Amino acid metabolism > Arginine biosynthesis.

· Metabolism > Amino acid metabolism > Arginine and proline metabolism.

· Metabolism > Global and overview maps > Metabolic pathways.

· Organismal Systems > Endocrine system > Relaxin signaling pathway.

References

1). Liu H et al. Potential T2DM drug HMPA promotes short‐chain fatty acid production by improving carbon catabolite repression effect of gut microbiota. Br J Pharmacol 2020 Dec 7. (PubMed: 33284460) [IF=9.473]

Application: WB    Species: rat    Sample: intestinal

Figure 3.| HMPA treatment increased the abundance of butyric acid-producing bacteria in gut microbiota and inhibited the iNOS expression of intestinal tissue in HFD-STZ-induced T2DM rats.(E) iNOS expression of intestinal tissue in each group, as detected by Western blot.

2). Mai S et al. Oesophageal squamous cell carcinoma–associated IL‐33 rewires macrophage polarization towards M2 via activating ornithine decarboxylase. Cell Prolif 2021 Feb;54(2):e12960. (PubMed: 33305406) [IF=8.755]

Application: WB    Species: human    Sample: M2 macrophages

FIGURE 4|IL-33 promotes M2 macrophage polarization via ornithine decarboxylase (ODC) activation.E, iNOS and ARG1 protein levels were determined by Western blot in IL-33–induced M2 macrophage with or without knocking down ODC. *P < .05; **P < .01; ***P < .001. Bars indicate mean and SEM of triplicate experiments and show a representative experiment of at least 3 independent experiments performed for each panel

3). Zhang M et al. Human Tissue Kallikrein 1 Is Downregulated in Elderly Human Prostates and Possesses Potential In Vitro Antioxidative and Antifibrotic Effects in Rodent Prostates. Oxid Med Cell Longev 2021 Apr 30;2021:8877540. (PubMed: 34007408) [IF=7.310]

Application: WB    Species: rat    Sample: prostate cells

Figure 7:| The direct effects of BK on prostate cells.(e, f ) TGF-β1 and iNOS protein expression within WPMY-1 and RPrPF under 10 nM BK by Western blot. Each bar represents mean ± SD of 3 independent repeated experiment. §§P < 0:01 (WPMY-1 + BK vs. WPMY-1 + vehicle). $$P < 0:01 (RPrPF + BK vs. RPrPF + vehicle).

4). Lu H et al. Rosiglitazone Suppresses Renal Crystal Deposition by Ameliorating Tubular Injury Resulted from Oxidative Stress and Inflammatory Response via Promoting the Nrf2/HO-1 Pathway and Shifting Macrophage Polarization. Oxid Med Cell Longev 2021 Oct 14;2021:5527137. (PubMed: 34691355) [IF=7.310]

5). Fan X et al. Design, synthesis and bioactivity study of N-salicyloyl tryptamine derivatives as multifunctional agents for the treatment of neuroinflammation. Eur J Med Chem 2020 May 1;193:112217. (PubMed: 32182488) [IF=7.088]

Application: WB    Species: Mouse    Sample: BV2 and C6 cells

Fig. 2. Effects of compounds on protein expressions of iNOS, COX-2 in LPS-induced BV2 cell lines (A) and effects of compounds on protein expressions of iNOS in LPS-induced C6 cell lines (B). BV2 cell lines were exposed to the indicated compounds at concentrations of 10, 20 and 40 mM for 24 h in the presence of LPS (1 mg/mL). C6 cell lines were exposed to the indicated compounds at concentrations of 10, 20 and 40 mM for 24 h in the presence of LPS (10 mg/mL). The expression of iNOS and COX-2 was assayed and calculated by image J. Each error bar represents the mean ± SD of three independent experiments. #, p < 0.05 versus the control group; ##, p < 0.01 versus the control group; ###, p < 0.001 versus the control group. *, p < 0.05, **, p < 0.01 when compared to the LPS-treated cells. All data are the mean ± SD of at least three independent experiments.

6). Zhang J et al. MicroRNA-24 inhibits the oxidative stress induced by vascular injury by activating the Nrf2/Ho-1 signaling pathway. Atherosclerosis 2019 Sep 9;290:9-18 (PubMed: 31539718) [IF=6.847]

Application: WB    Species: rat    Sample: arteries

Fig. 4. |miR-24 inhibited oxidative stress induced by balloon injury in diabetic rats. Protein expression was measured in balloon-injured carotid arteries were treated with Ad-miR-24 or Ad-Scramble for 7 d.(A–E) Western blot analysis showing Nox 2, Nox 4, Enos, and Inos protein expression in Ad-miR-24 and Ad-Scramble treated arteries. Gapdh was used as internal control.

7). Wang L et al. Suppression of NOD-like receptor protein 3 inflammasome activation and macrophage M1 polarization by hederagenin contributes to attenuation of sepsis-induced acute lung injury in rats. Bioengineered 2022 Mar;13(3):7262-7276. (PubMed: 35266443) [IF=6.832]

8). Li Z et al. Melatonin therapy protects against renal injury before and after release of bilateral ureteral obstruction in rats. Life Sci 2019 May 14 (PubMed: 31100324) [IF=6.780]

9). Yang T et al. Berberine regulates macrophage polarization through IL-4-STAT6 signaling pathway in Helicobacter pylori-induced chronic atrophic gastritis. Life Sci 2021 Feb 1;266:118903. (PubMed: 33340526) [IF=6.780]

Application: WB    Species: Mice    Sample: RAW 264.7 cells

Fig. 6. Effects of BBR on the protein expression in RAW 264.7 cells. (A) WB images of IL-4, STAT6, GATA3, IRF-8 and NOS2; (B) Relative IL-4 protein level analyzed by WB; (C) Relative STAT6 protein level analyzed by WB; (D) Relative GATA3 protein level analyzed by WB; (E) Relative IRF-8 protein level analyzed by WB; (F) Relative NOS2 protein level analyzed by WB; ##P < 0.01 versus control group; **P < 0.01 versus H. pylori group. IL-4, interleukin 4; STAT6, signal transducer and activator of transcription 6; GATA3, GATA binding protein 3; IRF8, interferon regulatory factor 8; NOS2, nitric oxide synthase 2; Ctrl, control group.

10). Yuan P et al. Kaempferol alleviates calcium oxalate crystal-induced renal injury and crystal deposition via regulation of the AR/NOX2 signaling pathway. Phytomedicine 2021 Mar 27;86:153555. (PubMed: 33852977) [IF=6.656]

Application: IHC    Species: Human    Sample: HK-2 cells

Fig. 3. Kaempferol inhibited renal oxidative stress and inflammation in vivo. (a-e) NOX2, iNOS, NFκB-p65 and F4/80 stainings of paraffin embedded kidney sections were evaluated. (f) ROS level in renal tissue was detected by fluorescence microplate after loading with DCFH-DA. Moreover, (g) H2O2 expression, as well as (h) GSH, SOD and MDA levels were detected in mice renal tissues. (i-n) The mRNA levels of IL-1β, IL-6, TNF-α, IL-10, IL-4 and Arg1 in renal tissues were measured by RT-qPCR. (o) Group annotations were applied for all histograms. Data were expressed as the fold changes of the experimental group to the NC group, and were represented as means ± SD. * p < 0.05 vs. NC group; ** p < 0.05 as GA + Kae (25 mg/kg) vs. GA group, GA + Kae (50 mg/kg) vs. GA group, and GA + Kae (25 mg/kg) vs. GA + Kae (50 mg/kg) group, respectively.

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