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AB251432

Anti-iNOS antibody [EPR16630] - BSA and Azide free

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Rabbit Recombinant Monoclonal iNOS antibody. Carrier free. Suitable for IP, ELISA, WB and reacts with Mouse, Rat, Recombinant full length protein - Mouse samples.

View Alternative Names

Inosl, Nos2, Inducible NO synthase, Macrophage NOS, NOS type II, Peptidyl-cysteine S-nitrosylase NOS2, Inducible NOS, iNOS, MAC-NOS

5 Images
Immunoprecipitation - Anti-iNOS antibody [EPR16630] - BSA and Azide free (AB251432)
  • IP

Supplier Data

Immunoprecipitation - Anti-iNOS antibody [EPR16630] - BSA and Azide free (AB251432)

This data was developed using ab205529, the same antibody clone in a different buffer formulation.

iNOS was immunoprecipitated from 1mg of RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) treated with 0.1 mg/ml LPS for 6 hours whole cell extract with ab205529 at 1/80 dilution. Western blot was performed from the immunoprecipitate using ab205529 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : RAW 264.7 treated with 0.1 mg/ml LPS for 6 hours whole cell extract 10ug (Input).
Lane 2 : ab205529 IP in RAW 264.7 treated with 0.1 mg/ml LPS for 6 hours whole cell extract.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab205529 in RAW 264.7 treated with 0.1 mg/ml LPS for 6 hours whole cell extract.

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

Exposure time : 8 seconds.

All lanes:

Immunoprecipitation - Anti-iNOS antibody [EPR16630] (<a href='/en-us/products/primary-antibodies/inos-antibody-epr16630-ab205529'>ab205529</a>)

Predicted band size: 131 kDa

false

ELISA - Anti-iNOS antibody [EPR16630] - BSA and Azide free (AB251432)
  • ELISA

Supplier Data

ELISA - Anti-iNOS antibody [EPR16630] - BSA and Azide free (AB251432)

This data was developed using ab205529, the same antibody clone in a different buffer formulation.ELISA analysis of House mouse iNOS recombinant protein at 1000 ng/mL with ab205529. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.

Western blot - Anti-iNOS antibody [EPR16630] - BSA and Azide free (AB251432)
  • WB

Supplier Data

Western blot - Anti-iNOS antibody [EPR16630] - BSA and Azide free (AB251432)

This data was developed using ab205529, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-iNOS antibody [EPR16630] (<a href='/en-us/products/primary-antibodies/inos-antibody-epr16630-ab205529'>ab205529</a>) at 1/1000 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 (Mouse macrophage cells transformed with Abelson murine leukemia virus) treated with 0.1 µg/ml LPS for 6 hours whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 131 kDa

Observed band size: 131 kDa

true

Exposure time: 1s

Western blot - Anti-iNOS antibody [EPR16630] - BSA and Azide free (AB251432)
  • WB

Supplier Data

Western blot - Anti-iNOS antibody [EPR16630] - BSA and Azide free (AB251432)

This data was developed using ab205529, the same antibody clone in a different buffer formulation.

The molecular weight observed and the treatment method are consistent with what has been described in the literature (PMID : 12062366)

All lanes:

Western blot - Anti-iNOS antibody [EPR16630] (<a href='/en-us/products/primary-antibodies/inos-antibody-epr16630-ab205529'>ab205529</a>) 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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 131 kDa

false

Western blot - Anti-iNOS antibody [EPR16630] - BSA and Azide free (AB251432)
  • WB

Lab

Western blot - Anti-iNOS antibody [EPR16630] - BSA and Azide free (AB251432)

This data was developed using ab205529, the same antibody clone in a different buffer formulation. Blocking and dilution 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 [EPR16630] (<a href='/en-us/products/primary-antibodies/inos-antibody-epr16630-ab205529'>ab205529</a>) 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

Observed band size: 131 kDa

false

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR16630

Isotype

IgG

Carrier free

Yes

Reacts with

Rat, Mouse

Applications

ELISA, WB, IP

applications

Immunogen

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

Specificity

We were unable to detect a positive signal in inflammation‑induced THP‑1 cells. According to published studies (PMID:33553532, PMID: 24477906), human macrophages produce very little detectable iNOS and NO in response to inflammatory stimuli due to epigenetic regulation around the NOS2 transcription start site.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "ELISA" : {"fullname" : "ELISA", "shortname":"ELISA"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Mouse": { "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "", "IP-species-notes": "<p></p>", "ELISA-species-checked": "guaranteed", "ELISA-species-dilution-info": "", "ELISA-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>" }, "Rat": { "IP-species-checked": "guaranteed", "IP-species-dilution-info": "", "IP-species-notes": "", "ELISA-species-checked": "guaranteed", "ELISA-species-dilution-info": "", "ELISA-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>" }, "Recombinant full length protein - Mouse": { "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "ELISA-species-checked": "testedAndGuaranteed", "ELISA-species-dilution-info": "", "ELISA-species-notes": "<p></p>", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "" } } }
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Product details

ab251432 is the carrier-free version of ab205529.

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

What are the advantages of a recombinant monoclonal antibody?
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.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. 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.

Compatibility
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.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze
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Supplementary information

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

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.

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.
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Product protocols

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

Produces nitric oxide (NO) which is a messenger molecule with diverse functions throughout the body (PubMed : 7503239). In macrophages, NO mediates tumoricidal and bactericidal actions. Also has nitrosylase activity and mediates cysteine S-nitrosylation of cytoplasmic target proteins such PTGS2/COX2 (PubMed : 16373578). As component of the iNOS-S100A8/9 transnitrosylase complex involved in the selective inflammatory stimulus-dependent S-nitrosylation of GAPDH implicated in regulation of the GAIT complex activity and probably multiple targets including ANXA5, EZR, MSN and VIM (By similarity). Involved in inflammation, enhances the synthesis of pro-inflammatory mediators such as IL6 and IL8 (By similarity).
See full target information Nos2
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Product promise

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For full details, please see our Terms & Conditions

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