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Anti-iNOS antibody ab15323 is a rabbit polyclonal antibody that is used in iNOS western blotting and IHC. Suitable for mouse samples.

- Tried and trusted by researchers since 2004


Images

Western blot - Anti-iNOS antibody (AB15323), expandable thumbnail
  • Western blot - Anti-iNOS antibody (AB15323), expandable thumbnail
  • Western blot - Anti-iNOS antibody (AB15323), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody (AB15323), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody (AB15323), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.4
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA

Form

Liquid

Clonality

Polyclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PWB
Human
Predicted
Predicted
Mouse
Expected
Expected
Rat
Predicted
Predicted
Recombinant full length protein - Mouse
Not recommended
Expected

Expected
Expected

Species

Mouse

Dilution info

1/100

Notes

Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Predicted
Predicted

Species

Rat, Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Recombinant full length protein - Mouse

Dilution info

-

Notes

-

Expected
Expected

Species

Recombinant full length protein - Mouse

Dilution info

-

Notes

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.

Species

Mouse

Dilution info

-

Notes

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.

Predicted
Predicted

Species

Rat, Human

Dilution info

-

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:7531687, PubMed:7544004). 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 proinflammatory mediators such as IL6 and IL8 (PubMed:19688109).

Alternative names

Recommended products

Anti-iNOS antibody ab15323 is a rabbit polyclonal antibody that is used in iNOS western blotting and IHC. Suitable for mouse samples.

- Tried and trusted by researchers since 2004

Key facts

Isotype

IgG

Form

Liquid

Clonality

Polyclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Purification technique

Affinity purification Immunogen

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.

Concentration
Loading...

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.

This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.

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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In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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5 product images

  • Western blot - Anti-iNOS antibody (ab15323), expandable thumbnail

    Western blot - Anti-iNOS antibody (ab15323)

    This 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 (Anti-GAPDH antibody [6C5] - Loading Control 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.

    All lanes: Western blot - Anti-iNOS antibody (ab15323) at 1 µg/mL

    Lane 1: Raw264.7 at 20 µg

    Lane 2: Raw264.7 + LPS (1 ug per mL; 18 hours) at 20 µg

    Secondary

    All lanes: IR- goat anti-rabbit (green) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 131 kDa

  • Western blot - Anti-iNOS antibody (ab15323), expandable thumbnail

    Western blot - Anti-iNOS antibody (ab15323)

    All lanes: Western blot - Anti-iNOS antibody (ab15323) at 5 µg/mL

    Lane 1: Raw 264.7 whole cell lysate + PMA at 20 µg

    Lane 2: Raw 264.7 whole cell lysate + PMA + LPS + Brefeldin A at 20 µg

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 131 kDa

    Observed band size: 140 kDa

    Exposure time: 8min

  • Western blot - Anti-iNOS antibody (ab15323), expandable thumbnail

    Western blot - Anti-iNOS antibody (ab15323)

    Membrane was blocked in 2% milk for 1 hour at RT. Primary anitbody was incubated at 4°C overnight.

    All lanes: Western blot - Anti-iNOS antibody (ab15323) at 5 µg/mL

    Lane 1: Human recombinant iNOS protein at 0.1 µg

    Lane 2: Mouse recombinant iNOS protein at 0.1 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (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, 47 kDa, 68 kDa

    Exposure time: 1min

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody (ab15323), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody (ab15323)

    IHC image of iNOS staining in a section of formalin-fixed paraffin-embedded normal mouse lung performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab15323, 1/100 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody (ab15323), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody (ab15323)

    IHC image of iNOS staining in a section of formalin-fixed paraffin-embedded normal rat lung performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab15323, 1/100 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com