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Anti-iNOS antibody ab3523 is a rabbit polyclonal antibody that is used in iNOS western blotting, IHC and immunofluorescence. Suitable for human and mouse samples.

- Tried and trusted by researchers since 2003


Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody (AB3523), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody (AB3523), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody (AB3523), expandable thumbnail
  • Western blot - Anti-iNOS antibody (AB3523), expandable thumbnail
  • Western blot - Anti-iNOS antibody (AB3523), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

Preservative: 0.05% Sodium azide
Constituents: 99% PBS, 0.1% BSA

Form

Liquid

Clonality

Polyclonal

Immunogen

  • Synthetic Peptide within Mouse Nos2 aa 1-50. The exact immunogen used to generate this antibody is proprietary information. Database link P29477

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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PICC/IFWB
Human
Tested
Tested
Expected
Mouse
Expected
Tested
Tested

Tested
Tested

Species

Human

Dilution info

1/20

Notes

-

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/50

Notes

-

Species

Human

Dilution info

1/50

Notes

-

Tested
Tested

Species

Mouse

Dilution info

-

Notes

-

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

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Target data

Function

Produces nitric oxide (NO) which is a messenger molecule with diverse functions throughout the body (PubMed:7504305, PubMed:7531687, PubMed:7544004, PubMed:7682706). 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 (PubMed:25417112). Involved in inflammation, enhances the synthesis of pro-inflammatory mediators such as IL6 and IL8 (PubMed:19688109).

Alternative names

Recommended products

Anti-iNOS antibody ab3523 is a rabbit polyclonal antibody that is used in iNOS western blotting, IHC and immunofluorescence. Suitable for human and mouse samples.

- Tried and trusted by researchers since 2003

Key facts

Isotype

IgG

Form

Liquid

Clonality

Polyclonal

Immunogen
  • Synthetic Peptide within Mouse Nos2 aa 1-50. The exact immunogen used to generate this antibody is proprietary information. Database link P29477
Purification technique

Affinity purification Immunogen

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.

Concentration
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Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

Abcam is leading the way to address reproducibility in scientific research with our highly validated recombinant monoclonal and recombinant multiclonal antibodies. Search & select one of Abcam's thousands of recombinant alternatives to eliminate batch-variability and unnecessary animal use.

If you do not find a host species to meet your needs, our catalogue and custom Chimeric range provides scientists the specificity of Abcam's RabMAbs in the species backbone of your choice. Remember to also review our range of edited cell lines, proteins and biochemicals relevant to your target that may help you further your research goals.

Abcam antibodies are extensively validated in a wide range of species and applications, so please check the reagent specifications meet your scientific needs before purchasing. If you have any questions or bespoke requirements, simply visit the Contact Us page to send us an inquiry or contact our Support Team ahead of purchase.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Inducible Nitric Oxide Synthase commonly referred to as iNOS or NOS2 is an important enzyme in the production of nitric oxide (NO) from L-arginine. This enzyme has a molecular weight of approximately 130 kDa and is expressed in various cell types including macrophages and hepatocytes. iNOS becomes active in response to inflammatory stimuli and can produce large amounts of NO for extended periods. Western blotting and iNOS ELISA kits are frequently used methods to detect and quantify the presence and activity of iNOS in different biological samples.

Biological function summary

INOS plays a significant role in the immune system by participating in the defense against pathogens. The enzyme is part of a molecular complex that generates NO which acts as a signaling molecule and has antimicrobial properties. iNOS activity is induced during immune responses and contributes to the elimination of bacteria and viruses through the production of reactive nitrogen species. Its expression is tightly regulated by cytokines and other signaling molecules that modulate the extent of its action.

Pathways

INOS integrates into the nitric oxide signaling pathway and is closely associated with inflammatory pathways. NO serves as a messenger within the nitric oxide signaling pathway facilitating processes like vasodilation and neurotransmission. iNOS works alongside other proteins such as cytokines like interferon-gamma that elevate its activity during inflammation and immune responses. The regulation of iNOS expression and activity plays an essential role in maintaining immune homeostasis and ensuring proper response to infections.

Associated diseases and disorders

INOS is linked to conditions characterized by excessive inflammation such as rheumatoid arthritis and sepsis. In these diseases overproduction of nitric oxide by iNOS can lead to tissue damage and exacerbate symptoms. Additionally iNOS is associated with the protein TNF-alpha as its expression is often driven by inflammatory mediators during disease states. Understanding the role of iNOS in such conditions provides insights into potential therapeutic targets for managing inflammatory diseases.

Product promise

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5 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody (ab3523), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody (ab3523)

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody (ab3523), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody (ab3523)

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody (ab3523), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody (ab3523)

    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.

  • Western blot - Anti-iNOS antibody (ab3523), expandable thumbnail

    Western blot - Anti-iNOS antibody (ab3523)

    Using 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.

    All lanes: Western blot - Anti-iNOS antibody (ab3523)

    Lane 1: RAW264 whole cell lysate untreated at 20 µg

    Lane 2: RAW264 whole cell lysate untreated stimulated with LPS at 1 µg/mL for 16 hours at 20 µg

    Secondary

    All lanes: Goat anti-Rabbit IgG (Heavy Chain) Superclonal Secondary Antibody, HRP conjugate at 1/1000 dilution

  • Western blot - Anti-iNOS antibody (ab3523), expandable thumbnail

    Western blot - Anti-iNOS antibody (ab3523)

    This 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.

    All lanes: Western blot - Anti-iNOS antibody (ab3523)

    Lane 1: Whole cell lysate of RAW 264.7 at 30 µg

    Lane 2: Whole cell lysate of RAW 264.7 treated with LPS at 30 µg

    Secondary

    All lanes: Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP

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Product protocols

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