Anti-iNOS antibody (ab15323)
Overview
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Product name
Anti-iNOS antibody
See all iNOS primary antibodies -
Description
Rabbit polyclonal to iNOS -
Host species
Rabbit -
Specificity
The immunogen used to raise this antibody shares 84% homology with Rat iNOS. Ab15323 has been batch tested using mouse lysates/tissues only. Some customers have successfully used ab15323 with rat. Please contact Abcam Scientific Support for more information.
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Tested applications
Suitable for: IHC-Fr, Electron Microscopy, WB, Flow Cyt, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat -
Immunogen
Synthetic peptide corresponding to Mouse iNOS.
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Positive control
- IHC-P: Mouse colon and spleen tissue; Rat liver and spleen tissue. WB: RAW 264.7 cells. Human and mouse recombinant iNOS protein. ICC/IF: NIH-3T3 cells. Flow Cyt: Mouse whole blood cells.
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General notes
This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.4
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
Our Abpromise guarantee covers the use of ab15323 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| IHC-Fr | Use at an assay dependent concentration. PubMed: 18523253 | |
| Electron Microscopy | Use at an assay dependent concentration. PubMed: 21071693 | |
| WB | Use at an assay dependent concentration. Detects a band of approximately 140 kDa (predicted molecular weight: 131 kDa). Please use at a assay dependent concentration. 1µg/ml was used of a 0.2mg/ml stock (lot GR126616-1). Otherwise 1/250 is recommended for any batches at lower concentration. |
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| Flow Cyt | Use at an assay dependent concentration. | |
| IHC-P | 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. | |
| ICC/IF | Use at an assay dependent concentration. PubMed: 20584290 |
Target
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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 COX2. -
Tissue specificity
Expressed in the liver, retina, bone cells and airway epithelial cells of the lung. Not expressed in the platelets. -
Sequence similarities
Belongs to the NOS family.
Contains 1 FAD-binding FR-type domain.
Contains 1 flavodoxin-like domain. - Information by UniProt
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Database links
- Entrez Gene: 18126 Mouse
- Entrez Gene: 24599 Rat
- SwissProt: P29477 Mouse
- SwissProt: Q06518 Rat
- Unigene: 2893 Mouse
- Unigene: 10400 Rat
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Alternative names
- HEP-NOS antibody
- Hepatocyte NOS antibody
- HEPNOS antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody (ab15323)Gentile D. et al PLoS One. 2018 Apr 11;13(4):e0195502. doi: 10.1371/journal.pone.0195502. eCollection 2018. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/Apigenin reduced SP and iNOS expression in colonic myenteric ganglia from HFD mice.
Representative pictures of SP (substance P) and iNOS immunostaining of cross-sections from full-thickness mice colonic specimens from SD, SD plus treatment with apigenin (10 mg/Kg/day), HFD or HFD plus treatment with apigenin (10 mg/Kg/day). Scale bar = 20 μm.
Paraffin-sections were processed for immunoperoxidase staining. After an overnight incubation with primary antibodies against SP (code n. Sc-21715, Santa Cruz Biotech, California, USA) and iNOS (code n. ab 15323, Abcam, Cambridge, UK), the slices were exposed to appropriate biotinylated immunoglobulins, peroxidase-labelled streptavidin complex, and 3.3’-diaminobenzidine tetrahydrochloride (DakoCytomation, Glostrup, Denmark). The quantitative assessment of the immunostained sections was performed by the L.A.S. software v.4.5 Image Analysis System. The expression of antigens was evaluated as percentage of positive pixels (PPP) estimated over the total area of myenteric ganglia examined.
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Western blot - Anti-iNOS antibody (ab15323)All lanes : Anti-iNOS antibody (ab15323) at 1 µg/ml
Lane 1 : Raw264.7
Lane 2 : Raw264.7 + LPS (1 ug per mL; 18 hours)
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : IR- goat anti-rabbit (green) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 131 kDaThis blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system under denaturing, reducing conditions. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with rabbit polyclonal to iNOS (ab15323; 1 ug x mL-1 (based on antibody concentration of 0.2mg/ml - lot GR126616-1) and the loading control mouse anti-GAPDH antibody (ab8245; 1:10000) overnight at 4°C. Antibody binding was detected using infrared (IR) labelled goat anti-rabbit (green; 1:10000) and IR-goat anti-mouse (red; 1:10000) for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody (ab15323)This image is courtesy of an Abreview submitted by Carl Hobbsab15323 staining rat liver (lower image) and spleen (upper image) sections by IHC-P. The tissue was fixed with formaldehyde and a heat mediated antigen retrival step was performed with citric acid pH 6. Blocking of the sample was done with 1%BSA for 10 minutes at 21°C, followed by staining with ab15323 at 1/100 in TBS/BSA/azide for 16h at 21°C. A biotinylated goat anti-rabbit polyclonal antibody at 1/200 was used as the secondary antibody.
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Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody (ab15323)Image is courtesy of an anonymous Abreviewab15323 staining iNOS in Mouse NIH3T3 cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton and blocked with 10% serum for 30 minutes at 24°C. Samples were incubated with primary antibody (1/100) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated Donkey anti-rabbit polyclonal was used as the secondary antibody (1/500).
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Flow Cytometry - Anti-iNOS antibody (ab15323)This image is courtesy of an anonymous Abreviewab15323 staining iNOS in Mouse whole blood cells by Flow Cytometry. The sample was incubated with the undiluted primary antibody for 30 minutes at 25°C. ab97050, FITC-conjugated Goat anti-rabbit IgG (undiluted) was used as the secondary antibody.
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Western blot - Anti-iNOS antibody (ab15323)All lanes : Anti-iNOS antibody (ab15323) at 5 µg/ml
Lane 1 : Raw 264.7 whole cell lysate + PMA
Lane 2 : Raw 264.7 whole cell lysate + PMA + LPS + Brefeldin A
Lysates/proteins at 20 µg per lane.
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 131 kDa
Additional bands at: 140 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 8 minutes -
Western blot - Anti-iNOS antibody (ab15323)All lanes : Anti-iNOS antibody (ab15323) at 5 µg/ml
Lane 1 : Human recombinant iNOS protein
Lane 2 : Mouse recombinant iNOS protein
Lysates/proteins at 0.1 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 131 kDa
Exposure time: 1 minuteMembrane was blocked in 2% milk for 1 hour at RT. Primary anitbody was incubated at 4°C overnight.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody (ab15323)This image is courtesy of an Abreview submitted by Carl Hobbsab15323 staining macrophages in mouse spleen sections by IHC-P. The tissue was fixed with formaldehyde and a heat mediated antigen retrival step was performed with citric acid pH 6. Blocking of the sample was done with 1%BSA for 10 minutes at 21°C, followed by staining with ab15323 at 1/50 in TBS/BSA/azide for 16h at 21°C. A biotinylated goat anti-rabbit polyclonal antibody at 1/200 was used as the secondary antibody.
Protocols
References
ab15323 has been referenced in 230 publications.
- Bloomer SA et al. Aging results in accumulation of M1 and M2 hepatic macrophages and a differential response to gadolinium chloride. Histochem Cell Biol 153:37-48 (2020). IHC-P ; Rat . PubMed: 31691025
- Wichaiyo S et al. Platelet glycoprotein VI and C-type lectin-like receptor 2 deficiency accelerates wound healing by impairing vascular integrity in mice. Haematologica 104:1648-1660 (2019). PubMed: 30733265
- Li HY et al. Lycium barbarum (Wolfberry) Increases Retinal Ganglion Cell Survival and Affects both Microglia/Macrophage Polarization and Autophagy after Rat Partial Optic Nerve Transection. Cell Transplant 28:607-618 (2019). PubMed: 30838886
- Liu M et al. C1q/TNF-related protein-9 promotes macrophage polarization and improves cardiac dysfunction after myocardial infarction. J Cell Physiol N/A:N/A (2019). PubMed: 30953351
- Zhou X et al. YAP Aggravates Inflammatory Bowel Disease by Regulating M1/M2 Macrophage Polarization and Gut Microbial Homeostasis. Cell Rep 27:1176-1189.e5 (2019). PubMed: 31018132
