Anti-iNOS antibody [RM1017] 20 ul size (ab283655)

Anti-iNOS antibody [RM1017] is a rabbit recombinant monoclonal antibody that is sed to detect inducible nitric oxide synthase (iNOS) in ICC/IF, western blotting, IHC-P, IP, and Flow cytometryometry (intra). Suitable for human, mouse, rat samples.

- Cited in over 25 publications

Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody [RM1017] (AB283655), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Multiclonal

Immunogen

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

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IHC-P	IP	Flow Cyt (Intra)	WB	I-ELISA	ICC/IF	IHC-Fr
Human	Tested	Tested	Tested	Tested	Expected	Not recommended	Not recommended
Mouse	Not recommended	Tested	Tested	Tested	Expected	Tested	Not recommended
Rat	Not recommended	Expected	Expected	Tested	Expected	Expected	Not recommended
Recombinant fragment - Human	Not recommended	Not recommended	Not recommended	Not recommended	Tested	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/2000	Notes We have preliminary internal testing data to indicate this antibody showed no staining in mouse and rat tissue samples by IHC-P. Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes We have preliminary internal testing data to indicate this antibody showed no staining in mouse and rat tissue samples by IHC-P.
Species Mouse	Dilution info -	Notes We have preliminary internal testing data to indicate this antibody showed no staining in mouse and rat tissue samples by IHC-P.
Species Recombinant fragment - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/30	Notes -
Species Human	Dilution info 1/30	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant fragment - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/600	Notes -
Species Human	Dilution info 1/600	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant fragment - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/2000	Notes -
Species Rat	Dilution info 1/2000	Notes -
Species Human	Dilution info 1/2000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant fragment - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Recombinant fragment - Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/50	Notes We have preliminary internal testing data to indicate this antibody may not be suitable for human ICC/IF

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes We have preliminary internal testing data to indicate this antibody may not be suitable for human ICC/IF
Species Recombinant fragment - Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Human, Recombinant fragment - Human	Dilution info -	Notes -

Target data

See full target information NOS2

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).

Additional Targets

Nos2

Alternative names

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

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Multiclonal

Immunogens

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

Clone number

RM1017

Purification technique

Affinity purification Protein A

Specificity

We have preliminary internal testing data to indicate this antibody may not be suitable for human ICC/IF and showed no staining in mouse and rat tissue samples by IHC-P.

FURTHER INFORMATION ON SPECIFICITY (Chinese Version) available under the support & downloads section.

Concentration

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

Product Specifications
Anti-iNOS antibody [RM1017] (ab283655) is a recombinant rabbit multiclonal antibody which is comprised of multiple monoclonal antibodies targeting different epitopes and recapitulating the performance of a polyclonal. Each component clone was developed by Abcam using patented rabbit monoclonal antibody technology.
Anti-iNOS antibody [RM1017] (ab283655) is validated for use in Flow Cyt (Intra), I-ELISA, ICC/IF, IHC-P, IP and WB in human, mouse and rat samples.
Anti-iNOS antibody [RM1017] (ab283655) specifically detects iNOS (UniProt ID: P35228; Molecular weight: 131kDa) and is sold in a convenient trial size to enable initial testing (20 µL) and larger sizes for subsequent scaling up experiments (100 µL and 1 mL).

Quality and Validation
Abcams high quality manufacturing and validation processes ensure Anti-iNOS antibody [RM1017] (ab283655) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.

Related Products
Conjugation-ready, carrier free format available for antibody clone RM1017 - ab283668.

What are recombinant multiclonals?
Recombinant multiclonals are a mixture of recombinant antibodies co-expressed from a library of heavy and light chains. They offer several advantages including:

- The sensitivity of polyclonal antibodies by recognising multiple epitopes
- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

View our range of recombinant multiclonal antibodies.

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.

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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11 product images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody [RM1017] (ab283655)
Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling iNOS with ab283655 at 1/2000 (0.3 μg/ml) followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection) was used. Positive staining on the hepatocytes in human liver. The section was incubated with ab283655 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
Immunoprecipitation - Anti-iNOS antibody [RM1017] (ab283655)
iNOS was immunoprecipitated from 0.35 mg RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 1μg/ml LPS for 6h whole cell lysate 10μg with ab283655 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283655 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 1ug/ml LPS for 6h whole cell lysate 10μg
Lane 2: ab283655 IP in RAW 264.7 treated with 1μg/ml LPS for 6h whole cell lysate
Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab283655 in RAW 264.7 treated with 1μg/ml LPS for 6h whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 8 seconds
Lower bands could be proteolytic cleavage as described in literature. (PMID: 11786228)
All lanes: Immunoprecipitation - Anti-iNOS antibody [RM1017] (ab283655)
Predicted band size: 131 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody [RM1017] (ab283655)
Immunohistochemical analysis of paraffin-embedded Human lung tissue labeling iNOS with ab283655 at 1/2000 (0.3 μg/ml) followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection) was used. Positive staining on the human lung. The section was incubated with ab283655 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
Flow Cytometry (Intracellular) - Anti-iNOS antibody [RM1017] (ab283655)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Caco-2 (Human colorectal adenocarcinoma epithelial cell) cells labelling iNOS with ab283655 at 1/600 dilution (0.1μg) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-iNOS antibody [RM1017] (ab283655)
Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labeling iNOS with ab283655 at 1/2000 (0.3 μg/ml) followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection) was used. Positive staining on the human lung carcinoma. The section was incubated with ab283655 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins
Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody [RM1017] (ab283655)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 cells labelling iNOS with ab283655 at 1/50 (12.0 μg/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 μg/ml) dilution (Green). Confocal image showing cytoplasmic staining in RAW 264.7 cell line after treatment with lipopolysaccharide ( 1ug/ml ) for 6 h. is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 (2.5μg/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 μg/ml) dilution.
Flow Cytometry (Intracellular) - Anti-iNOS antibody [RM1017] (ab283655)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 1ug/ml LPS for 6h (Red) / Untreated control (Green) cells labelling iNOS with ab283655 at 1/600 dilution (0.1ug) (Red) and Green (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody.
Indirect ELISA - Anti-iNOS antibody [RM1017] (ab283655)
Indirect ELISA using ab283655 at varying antibody concentrations and antigen concentration at 1000 ng/mL. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) (1/2500) was used as the secondary antibody.
Western blot - Anti-iNOS antibody [RM1017] (ab283655)
Blocking buffer and concentration: 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).
Lanes 1 - 3: Western blot - Anti-iNOS antibody [RM1017] (ab283655) at 1/1000 dilution
Lanes 1 - 3: Western blot - Anti-iNOS antibody [RM1017] - BSA and Azide free (Anti-iNOS antibody [RM1017] - BSA and Azide free ab283668)
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
Immunoprecipitation - Anti-iNOS antibody [RM1017] (ab283655)
iNOS was immunoprecipitated from DLD-1 (human colorectal adenocarcinoma epithelial cell) treated with 100U/mL IFN gamma (human), 50U/mL IL-1 beta (human) and 10ng/mL TNF alpha for 24h whole cell lysate with ab283655 at 1/30 dilution (2 μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283655 at 1/2000 dilution. VeriBlot for IP secondary antibody(HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: DLD-1 (human colorectal adenocarcinoma epithelial cell) treated with 100U/mL IFN gamma(human), 50U/mL IL-1 beta(human) and 10ng/mL TNF alpha for 24h whole cell lysate

Lane 2: Anti-YB1 antibody [RM1188] ab320005 IP in DLD-1 (human colorectal adenocarcinoma epithelial cell) treated with 100U/mL IFN gamma(human), 50U/mL IL-1 beta(human) and 10ng/mL TNF alpha for 24h whole cell lysate

Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab283655 in DLD-1 treated with 100U/mL IFN gamma (human), 50U/mL IL-1 beta (human) and 10ng/mL TNF alpha for 24h whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST
All lanes: Immunoprecipitation - Anti-iNOS antibody [RM1017] (ab283655) at 1/2000 dilution
All lanes: DLD-1 (human colorectal adenocarcinoma epithelial cell) treated with 100U/mL IFN gamma(human), 50U/mL IL-1 beta(human) and 10ng/mL TNF alpha for 24h whole cell lysate at 10 µg
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Predicted band size: 131 kDa
Observed band size: 131 kDa
Exposure time: 32s
Western blot - Anti-iNOS antibody [RM1017] (ab283655)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-iNOS antibody [RM1017] (ab283655) at 1/2000 dilution
Lane 1: DLD-1 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: DLD-1 treated with 100U/mL IFN gamma(human), 50U/mL IL-1 beta(human) and 10ng/mL TNF alpha for 24h whole cell lysate at 20 µg
Lane 3: Caco-2 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 4: RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg
Lane 5: RAW 264.7 treated with 100ng/ml lipopolysaccharide (LPS) for 6h whole cell lysate at 20 µg
Lane 6: NR8383 (rat alveolar macrophage) whole cell lysate at 20 µg
Lane 7: NR8383 treated with 1ng/ml lipopolysaccharide (LPS) for 24h 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 1/100000 dilution
Observed band size: 131 kDa, 36 kDa