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
IgG
Rabbit
pH: 7.4
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA
Liquid
Polyclonal
IHC-P | WB | |
---|---|---|
Human | Predicted | Predicted |
Mouse | Expected | Expected |
Rat | Predicted | Predicted |
Recombinant full length protein - Mouse | Not recommended | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/100 | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Recombinant full length protein - Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
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. |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Human | Dilution info - | Notes - |
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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).
Hepatocyte NOS, Inducible NO synthase, NOS type II, Peptidyl-cysteine S-nitrosylase NOS2, HEP-NOS, Inducible NOS, iNOS, NOS2, NOS2A
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
IgG
Rabbit
pH: 7.4
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA
Liquid
Polyclonal
Affinity purification Immunogen
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.
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
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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.
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.
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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This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
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Full details and terms and conditions can be found here:
Terms & Conditions.
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
All lanes: IR- goat anti-rabbit (green) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 131 kDa
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
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
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
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.
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.
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