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Anti-iNOS antibody [EPR16635] ab178945 is a rabbit monoclonal antibody that is used in iNOS western blotting and immunofluorescence. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone EPR16635 is cited in over 450 publications


Images

Western blot - Anti-iNOS antibody [EPR16635] (AB178945), expandable thumbnail
  • Immunoprecipitation - Anti-iNOS antibody [EPR16635] (AB178945), expandable thumbnail
  • Western blot - Anti-iNOS antibody [EPR16635] (AB178945), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody [EPR16635] (AB178945), expandable thumbnail
  • Western blot - Anti-iNOS antibody [EPR16635] (AB178945), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
I-ELISAWBICC/IFIP
Human
Expected
Tested
Expected
Expected
Mouse
Expected
Tested
Tested
Tested
Rat
Expected
Tested
Expected
Expected
Recombinant full length protein - Mouse
Tested
Not recommended
Not recommended
Not recommended

Tested
Tested

Species

Recombinant full length protein - Mouse

Dilution info

-

Notes

-

Expected
Expected

Species

Mouse, Rat, Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/1000

Notes

To detect iNOS expression, macrophages need to be treated with LPS (0.1 - 1 ug/ml).

Species

Rat

Dilution info

1/1000

Notes

To detect iNOS expression, macrophages need to be treated with LPS (0.1 - 1 ug/ml).

Species

Human

Dilution info

1/1000

Notes

To detect iNOS expression, macrophages need to be treated with LPS (0.1 - 1 ug/ml).

Not recommended
Not recommended

Species

Recombinant full length protein - Mouse

Dilution info

-

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/500

Notes

To detect iNOS expression, macrophages need to be treated with LPS (0.1 - 1 ug/ml).

Expected
Expected

Species

Rat, Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Recombinant full length protein - Mouse

Dilution info

-

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/100

Notes

To detect iNOS expression, macrophages need to be treated with LPS (0.1 - 1 ug/ml).

Expected
Expected

Species

Rat, Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Recombinant full length protein - Mouse

Dilution info

-

Notes

-

Associated Products

Select an associated product type

9 products for Alternative Product

1 product for Alternative Version

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

Hepatocyte NOS, Inducible NO synthase, NOS type II, Peptidyl-cysteine S-nitrosylase NOS2, HEP-NOS, Inducible NOS, iNOS, NOS2, NOS2A

Recommended products

Anti-iNOS antibody [EPR16635] ab178945 is a rabbit monoclonal antibody that is used in iNOS western blotting and immunofluorescence. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone EPR16635 is cited in over 450 publications

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

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

EPR16635

Purification technique

Affinity purification Protein A

Specificity

This antibody shows low affinity on human samples.
Based on our preliminary data, this antibody is not suitable for THP1 (Human monocytic leukemia monocyte) cell lines in WB.

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

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

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

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

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:
Terms & Conditions.

6 product images

  • Western blot - Anti-iNOS antibody [EPR16635] (ab178945), expandable thumbnail

    Western blot - Anti-iNOS antibody [EPR16635] (ab178945)

    Lanes 1 - 2: Western blot - Anti-iNOS antibody [EPR16635] – Mouse IgG2b (Chimeric) (Anti-iNOS antibody [EPR16635] - Mouse IgG2b (Chimeric) ab210823) at 1/1000 dilution

    Lanes 1 - 2: Western blot - Anti-iNOS antibody [EPR16635] (ab178945) at 1/1000 dilution

    Lane 1: Untreated L6 (rat skeletal muscle myoblast) whole cell lysate at 20 µg

    Lane 2: L6 treated with 50 ng/ml IL-1 beta, 20 ng/ml TNF-alpha and 100U/ml IFN-gamma for 24 h, whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 131 kDa

    Observed band size: 131 kDa

    This data was developed using ab178945, the same antibody clone in the original rabbit isotype.

  • Immunoprecipitation - Anti-iNOS antibody [EPR16635] (ab178945), expandable thumbnail

    Immunoprecipitation - Anti-iNOS antibody [EPR16635] (ab178945)

    iNOS was immunoprecipitated from 1mg of RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate treated with 1 μg/ml LPS for 24h with ab178945 at 1/100 dilution.

    Lane 1: RAW 264.7 whole cell lysate treated with 1 μg/ml LPS for 24h,10ug (Input).

    Lane 2: ab178945 IP in RAW 264.7 whole cell lysate treated with 1 μg/ml LPS for 24h.

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab178945 in RAW 264.7 whole cell lysate treated with 1 μg/ml LPS for 24h.

    Western blot was performed from the immunoprecipitate using ab178945 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used for detection at 1/1500.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 30 seconds.

    All lanes: Immunoprecipitation - Anti-iNOS antibody [EPR16635] (ab178945)

    Predicted band size: 131 kDa

  • Western blot - Anti-iNOS antibody [EPR16635] (ab178945), expandable thumbnail

    Western blot - Anti-iNOS antibody [EPR16635] (ab178945)

    Blocking and dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-iNOS antibody [EPR16635] (ab178945) at 1/20000 dilution

    Lane 1: Untreated RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate at 20 µg

    Lane 2: RAW 264.7 whole cell lysate treated with 0.1 µg/mL LPS for 6 hours at 20 µg

    Secondary

    All lanes: Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 131 kDa

    Observed band size: 131 kDa

    Exposure time: 30s

  • Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody [EPR16635] (ab178945), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody [EPR16635] (ab178945)

    Immunocytochemistry/Immunofluorescence analysis of RAW 264.7 non-treated and LPS treated (0.1 μg/mL) cells labelling iNOS with ab178945 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. The cells were co-stained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889, Alexa Fluor® 594 conjugated anti-alpha tubulin (1/200). Nuclei counterstained with DAPI (blue).

    Secondary antibody only controls performed on non-treated and treated cells.

  • Western blot - Anti-iNOS antibody [EPR16635] (ab178945), expandable thumbnail

    Western blot - Anti-iNOS antibody [EPR16635] (ab178945)

    All lanes: Western blot - Anti-iNOS antibody [EPR16635] (ab178945) at 1/500 dilution

    Lane 1: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 15 µg

    Lane 2: HepG2 (Human hepatocellular carcinoma epithelial cell) treated with 10ug/ml lipopolysaccharides for 6 hours whole cell lysates at 15 µg

    Lane 3: THP-1 (Human monocytic leukemia monocyte) whole cell lysates at 15 µg

    Lane 4: THP-1 (Human monocytic leukemia monocyte) treated with 100ng/ml lipopolysaccharides for 3 hours whole cell lysates at 15 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/200000 dilution

    Predicted band size: 131 kDa

    Exposure time: 3min

  • Western blot - Anti-iNOS antibody [EPR16635] (ab178945), expandable thumbnail

    Western blot - Anti-iNOS antibody [EPR16635] (ab178945)

    Blocking and diluting buffer 5% NFDM/TBST.

    iNOS is not normally expressed in the brain, but can be induced in the brain after inflammatory, infectious, or other damages (PMID: 11138926, PMID: 16156895, PMID: 10322315).

    All lanes: Western blot - Anti-iNOS antibody [EPR16635] (ab178945) at 1/1000 dilution

    Lane 1: Mouse hippocampus tissue lysate at 20 µg

    Lane 2: Mouse colon tissue lysate at 20 µg

    Lane 3: Mouse colon cancer tissue lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Observed band size: 131 kDa

    Exposure time: 180s

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Product protocols

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

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