Overview

  • Product nameAnti-iNOS antibody
    See all iNOS primary antibodies
  • Description
    Rabbit polyclonal to iNOS
  • SpecificityThis antibody detects iNOS. It does not detect other NOS isoforms. By Western blot, this antibody detects an ~135 kDa protein representing recombinant Human iNOS and Human iNOS from cytokine stimulated A549 cells. By Western blot, this antibody also detects purified recombinant Mouse iNOS, Mouse iNOS from cytokine stimulated RAW 264.7 cells.
  • Tested applicationsSuitable for: WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Chicken, Dog, Human, Xenopus laevis, Amphibians, Xenopus tropicalis
    Predicted to work with: Guinea pig
  • Immunogen

    Synthetic peptide corresponding to Mouse iNOS aa 17-31.
    Sequence:

    DLKEEKDINNNVKKT

  • Positive control
    • WB: recombinant Human or Mouse iNOS, cytokine stimulated A549 cell lysate, cytokine stimulated RAW 264.7 cell lysate, cytokine stimulated rat fibroblast lysate, LPS treated Mouse spleen

Properties

Applications

Our Abpromise guarantee covers the use of ab3523 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/200 - 1/800. Detects a band of approximately 135 kDa.

Abcam recommends using 5% BSA as the blocking agent. 

Note: Regarding using this antibody in WB on mouse lung tissue: "However, iNOS protein was below the limit of detection following any level of hypoxic exposure (positive control was lipopolysaccharide-treated mouse spleen) and using several different antibodies" (from PubMed ID 11686901)

Target

  • FunctionProduces nitric oxide (NO) which is a messenger molecule with diverse functions throughout the body. In macrophages, NO mediates tumoricidal and bactericidal actions. Also has nitrosylase activity and mediates cysteine S-nitrosylation of cytoplasmic target proteins such COX2.
  • Tissue specificityExpressed in the liver, retina, bone cells and airway epithelial cells of the lung. Not expressed in the platelets.
  • Sequence similaritiesBelongs to the NOS family.
    Contains 1 FAD-binding FR-type domain.
    Contains 1 flavodoxin-like domain.
  • Information by UniProt
  • Database links
  • Alternative names
    • HEP-NOS antibody
    • Hepatocyte NOS antibody
    • HEPNOS antibody
    • inducible antibody
    • Inducible nitric oxide synthase antibody
    • Inducible NO synthase antibody
    • Inducible NOS antibody
    • iNOS antibody
    • MAC NOS antibody
    • Macrophage NOS antibody
    • Nitric oxide synthase 2 inducible antibody
    • Nitric oxide synthase 2 inducible macrophage antibody
    • nitric oxide synthase 2A (inducible, hepatocytes) antibody
    • Nitric oxide synthase antibody
    • Nitric oxide synthase inducible antibody
    • nitric oxide synthase, macrophage antibody
    • NOS 2 antibody
    • NOS antibody
    • Nos II antibody
    • NOS type II antibody
    • nos2 antibody
    • NOS2_HUMAN antibody
    • NOS2A antibody
    • NOS2A, Inducible, Hepatocyte antibody
    • Peptidyl-cysteine S-nitrosylase NOS2 antibody
    see all

Anti-iNOS antibody images

  • All lanes : Anti-iNOS antibody (ab3523) at 1/500 dilution

    Lane 1 : iNOS Overexpression Lysate (Human)
    Lane 2 : Raw264.7 Whole Cell Lysate
    Lane 3 : Raw264.7 Whole Cell Lysate (LPS stimulated)

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat anti Rabbit IR680 at 1/10000 dilution

    Performed under reducing conditions.

    Observed band size : 140 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 50 kDa,70 kDa. We are unsure as to the identity of these extra bands.

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab3523 overnight at 4°C. Antibody binding was detected using Goat anti Rabbit IR680 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx

  • All lanes : Anti-iNOS antibody (ab3523) at 1/1000 dilution

    Lane 1 : Untreated RAW267.4 whole cell lysate.
    Lane 2 : RAW267.4 whole cell lysate treated with LPS at 1ug/ml for 16 hours.

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-rabbit IgG, HRP-linked at 1/2000 dilution
    developed using the ECL technique

    Exposure time : 5 minutes

    This image is curtesy of an anonymous AbReview.

    The cells were blocked with 5% milk for 2 hours at 22°C.

    See Abreview

References for Anti-iNOS antibody (ab3523)

This product has been referenced in:
  • Singh VP  et al. Metabolic Syndrome Is Associated with Increased Oxo-Nitrative Stress and Asthma-Like Changes in Lungs. PLoS One 10:e0129850 (2015). WB ; Mouse . Read more (PubMed: 26098111) »
  • Zhao J  et al. Synthetic Oligodeoxynucleotides Containing Multiple Telemeric TTAGGG Motifs Suppress Inflammasome Activity in Macrophages Subjected to Oxygen and Glucose Deprivation and Reduce Ischemic Brain Injury in Stroke-Prone Spontaneously Hypertensive Rats. PLoS One 10:e0140772 (2015). WB ; Rat . Read more (PubMed: 26473731) »

See all 24 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Liver)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric Acid
Permeabilization Yes - Triton X 100
Specification Liver
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative Formaldehyde
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Submitted May 21 2015

Thank you for contacting us.

Please send me your WB image so I can take a look at the bands you have obtained.

I look forward to hear back from you.

Application Western blot
Sample Rat Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions Reduced Denaturing (8%)
Loading amount 20 µg
Specification Skeletal muscle
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
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Submitted Mar 07 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions Reduced Denaturing (8%)
Loading amount 20 µg
Specification Skeletal muscle
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
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Submitted Mar 07 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Prostate Cancer)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate buffer pH 6.0
Permeabilization No
Specification Prostate Cancer
Blocking step Serum as blocking agent for 15 minute(s) · Concentration: 10% · Temperature: 23°C
Fixative Formaldehyde
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Submitted Nov 12 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (kidney)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate buffer 1X
Permeabilization No
Specification kidney
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative Formaldehyde
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Submitted Oct 14 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate pH6
Sample Pig Tissue sections (brain)
Specification brain
Permeabilization Yes - Triton
Fixative Paraformaldehyde
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Submitted Nov 25 2014

Application Western blot
Loading amount 20 µg
Gel Running Conditions Non-reduced Non-Denaturing (Native)
Sample Mouse Tissue lysate - whole (skeleton muscle, brain, bowel, lung, stomach)
Specification skeleton muscle, brain, bowel, lung, stomach
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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Submitted Jun 13 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Non-reduced Non-Denaturing (Native) (10)
Sample Rabbit Cell lysate - whole cell (microglia)
Specification microglia
Treatment 100ng/ml for 24hours
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
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Submitted Jun 12 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Rat Tissue sections (nerve)
Specification nerve
Fixative Paraformaldehyde
Permeabilization Yes - 0.2% triton
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
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Submitted May 08 2013

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"