Anti-iNOS antibody (ab3523)
Key features and details
- Rabbit polyclonal to iNOS
- Suitable for: IHC-P, WB, ICC/IF
- Reacts with: Mouse, Human
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
- Research with confidence – consistent and reproducible results with every batch
- Long-term and scalable supply – powered by recombinant technology for fast production
- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
Overview
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Product name
Anti-iNOS antibody
See all iNOS primary antibodies -
Description
Rabbit polyclonal to iNOS -
Host species
Rabbit -
Specificity
This antibody detects iNOS. It does not detect other NOS isoforms. By western blot, this antibody detects an ~135 kDa protein representing recombinant human iNOS. By western blot, this antibody also detects purified recombinant mouse iNOS, mouse iNOS from cytokine stimulated RAW 264.7 cells.
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Tested applications
Suitable for: IHC-P, WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Synthetic peptide corresponding to Mouse iNOS aa 1-100.
Database link: P29477 -
Positive control
- ICC: A549, NIH/3T3 cells. IHC-P: Human heart tissue.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 99% PBS -
Concentration information loading...
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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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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab3523 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 |
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IHC-P | (12) |
1/20.
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WB | (15) |
1/200 - 1/1000. Predicted molecular weight: 131 kDa.
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ICC/IF | (2) |
1/50.
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Notes |
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IHC-P
1/20. |
WB
1/200 - 1/1000. Predicted molecular weight: 131 kDa. |
ICC/IF
1/50. |
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: 4843 Human
- Entrez Gene: 18126 Mouse
- Omim: 163730 Human
- SwissProt: P35228 Human
- SwissProt: P29477 Mouse
- Unigene: 709191 Human
- Unigene: 2893 Mouse
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Alternative names
- HEP-NOS antibody
- Hepatocyte NOS antibody
- HEPNOS antibody
see all
Images
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All lanes : Anti-iNOS antibody (ab3523) at 1/1000 dilution
Lane 1 : RAW264 whole cell lysate untreated
Lane 2 : RAW264 whole cell lysate untreated stimulated with LPS at 1 µg/mL for 16 hours
Lysates/proteins at 20 µg per lane.
Secondary
Lane 1 : Goat anti-Rabbit IgG (Heavy Chain) Superclonal Secondary Antibody, HRP conjugate at 1/1000 dilution
Lane 2 : Goat anti-Rabbit IgG (Heavy Chain) HRP conjugate at 1/1000 dilution
Predicted band size: 131 kDaUsing 4-20% Tris-Glycine polyacrylamide gel and transferred to a nitrocellulose membrane, blocked with 5% Milk in TBST for at least 1 hour. The membrane was probed with ab3523 at 4°C overnight on a rocking platform, washed in TBST, and probed with the secondary antibody for 1 hour.
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All lanes : Anti-iNOS antibody (ab3523) at 1/1000 dilution
Lane 1 : Whole cell lysate of RAW 264.7
Lane 2 : Whole cell lysate of RAW 264.7 treated with LPS
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP at 1/4000 dilution
Predicted band size: 131 kDaThis was run using 4-12% Bis-Tris Protein Gel and a Nitrocellulose membrane.
Observed band at 130kDa in LPS treated RAW 264.7 cells, and an uncharacterized band at ~60kDa.
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Immunohistochemistry analysis of human heart tissue stained for iNOS without (negative control) or using ab3523 at 1/200 dilution overnight at 4°C in a humidified chamber, followed biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor.
To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature.
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Immunofluorescence analysis of A549 (human lung carcinoma cell line) whole cells labelling iNOS (Left panel: green) without (control) or using ab3523 at 1/20 dilution overnight at 4°C, followed DyLight-488 conjugated secondary antibody. Counter stain: F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.
Cells were grown on chamber slides and fixed with formaldehyde prior to staining.
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Immunofluorescence analysis of NIH/3T3 (mouse embryo fibroblast cell line) whole cells labelling iNOS (Left panel: green) without (control) or using ab3523 at 1/20 dilution overnight at 4°C, followed DyLight-488 conjugated secondary antibody. Counter stain: F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.
Cells were grown on chamber slides and fixed with formaldehyde prior to staining.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (313)
ab3523 has been referenced in 313 publications.
- Ruger L et al. Mechanical High-Intensity Focused Ultrasound (Histotripsy) in Dogs With Spontaneously Occurring Soft Tissue Sarcomas. IEEE Trans Biomed Eng 70:768-779 (2023). PubMed: 36006886
- Sallese A et al. Isolation and Characterization of Peritoneal Macrophages from Salamanders. Methods Mol Biol 2562:259-270 (2023). PubMed: 36272082
- Patino E et al. Iron therapy mitigates chronic kidney disease progression by regulating intracellular iron status of kidney macrophages. JCI Insight 8:N/A (2023). PubMed: 36394951
- Allouch A et al. CDKN1A is a target for phagocytosis-mediated cellular immunotherapy in acute leukemia. Nat Commun 13:6739 (2022). PubMed: 36347876
- Zhao S et al. GM-CSF-mediated inducement of bone marrow MDSCs by TSA and effect on survival of graft in mice. Eur J Med Res 27:161 (2022). PubMed: 36031660