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Rabbit Recombinant Monoclonal iNOS antibody. Carrier free. Suitable for ICC/IF, IP, WB, I-ELISA and reacts with Mouse, Rat, Human, Recombinant fragment - Mouse samples. Cited in 15 publications.

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Images

Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody [EPR16635] - BSA and Azide free (AB213987), expandable thumbnail
  • Immunoprecipitation - Anti-iNOS antibody [EPR16635] - BSA and Azide free (AB213987), expandable thumbnail
  • Indirect ELISA - Anti-iNOS antibody [EPR16635] - BSA and Azide free (AB213987), expandable thumbnail
  • Western blot - Anti-iNOS antibody [EPR16635] - BSA and Azide free (AB213987), expandable thumbnail
  • Western blot - Anti-iNOS antibody [EPR16635] - BSA and Azide free (AB213987), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: 100% PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ICC/IFIPWBI-ELISA
Human
Expected
Expected
Tested
Expected
Mouse
Tested
Tested
Tested
Expected
Rat
Expected
Expected
Tested
Expected
Recombinant fragment - Mouse
Not recommended
Not recommended
Not recommended
Tested

Tested
Tested

Species

Mouse

Dilution info

1/500

Notes

-

Expected
Expected

Species

Rat, Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Recombinant fragment - Mouse

Dilution info

-

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/100

Notes

-

Expected
Expected

Species

Rat, Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Recombinant fragment - Mouse

Dilution info

-

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/1000

Notes

-

Species

Rat

Dilution info

1/1000

Notes

-

Species

Human

Dilution info

1/1000

Notes

-

Not recommended
Not recommended

Species

Recombinant fragment - Mouse

Dilution info

-

Notes

-

Tested
Tested

Species

Recombinant fragment - Mouse

Dilution info

1000 ng/mL

Notes

-

Expected
Expected

Species

Mouse, Rat, Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

3 products for Alternative Version

2 products for Alternative Product

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

Rabbit Recombinant Monoclonal iNOS antibody. Carrier free. Suitable for ICC/IF, IP, WB, I-ELISA and reacts with Mouse, Rat, Human, Recombinant fragment - Mouse samples. Cited in 15 publications.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

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

Yes

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 conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

Notes

ab213987 is the carrier-free version of Anti-iNOS antibody [EPR16635] ab178945.

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.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

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.

7 product images

  • Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody [EPR16635] - BSA and Azide free (ab213987), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody [EPR16635] - BSA and Azide free (ab213987)

    Clone EPR16635 (ab213987) has been successfully conjugated by Abcam. This image was generated using Anti-iNOS antibody [EPR16635] (Alexa Fluor® 647). Please refer to Alexa Fluor® 647 Anti-iNOS antibody [EPR16635] ab209027 for protocol details.

    Alexa Fluor® 647 Anti-iNOS antibody [EPR16635] ab209027 staining iNOS in Raw264.7 cells. The lower panel shows cells treated with 1ug/ml Lipopolysaccharides (24 hr).  The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab209027at 1/100 dilution (shown in red) and Alexa Fluor® 488 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This product also gave a positive signal under the same testing conditions in SW480 cells fixed with 4% formaldehyde (10 min).

  • Immunoprecipitation - Anti-iNOS antibody [EPR16635] - BSA and Azide free (ab213987), expandable thumbnail

    Immunoprecipitation - Anti-iNOS antibody [EPR16635] - BSA and Azide free (ab213987)

    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 Anti-iNOS antibody [EPR16635] 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: Anti-iNOS antibody [EPR16635] 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 Anti-iNOS antibody [EPR16635] ab178945 in RAW 264.7 whole cell lysate treated with 1 μg/ml LPS for 24h.

    Western blot was performed from the immunoprecipitate using Anti-iNOS antibody [EPR16635] 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 dilution

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

    Exposure time: 30 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-iNOS antibody [EPR16635] ab178945).

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

    Predicted band size: 131 kDa

  • Indirect ELISA - Anti-iNOS antibody [EPR16635] - BSA and Azide free (ab213987), expandable thumbnail

    Indirect ELISA - Anti-iNOS antibody [EPR16635] - BSA and Azide free (ab213987)

    ELISA analysis of House mouse iNOS recombinant protein at 1000 ng/mL with Anti-iNOS antibody [EPR16635] ab178945. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-iNOS antibody [EPR16635] ab178945).

  • Western blot - Anti-iNOS antibody [EPR16635] - BSA and Azide free (ab213987), expandable thumbnail

    Western blot - Anti-iNOS antibody [EPR16635] - BSA and Azide free (ab213987)

    Blocking and dilution buffer: 5% NFDM/TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-iNOS antibody [EPR16635] ab178945).

    All lanes: Western blot - Anti-iNOS antibody [EPR16635] - BSA and Azide free (ab213987) 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

  • Western blot - Anti-iNOS antibody [EPR16635] - BSA and Azide free (ab213987), expandable thumbnail

    Western blot - Anti-iNOS antibody [EPR16635] - BSA and Azide free (ab213987)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-iNOS antibody [EPR16635] ab178945).

    All lanes: Western blot - Anti-iNOS antibody [EPR16635] - BSA and Azide free (ab213987) 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

  • Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody [EPR16635] - BSA and Azide free (ab213987), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody [EPR16635] - BSA and Azide free (ab213987)

    Immunocytochemistry/Immunofluorescence analysis of RAW 264.7 non-treated and LPS treated (0.1 μg/mL) cells labelling iNOS with Anti-iNOS antibody [EPR16635] 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-iNOS antibody [EPR16635] ab178945).

  • Western blot - Anti-iNOS antibody [EPR16635] - BSA and Azide free (ab213987), expandable thumbnail

    Western blot - Anti-iNOS antibody [EPR16635] - BSA and Azide free (ab213987)

    This data was developed using the same antibody clone in a different buffer formulation (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] (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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