Overview

  • Product name
    Anti-iNOS antibody
    See all iNOS primary antibodies
  • Description
    Rabbit polyclonal to iNOS
  • Host species
    Rabbit
  • Specificity
    This antibody detects iNOS. It does not detect other NOS isoforms. By western blot, this antibody detects an ~135 kDa protein representing recombinant human iNOS. By western blot, this antibody also detects purified recombinant mouse iNOS, mouse iNOS from cytokine stimulated RAW 264.7 cells.
  • Tested applications
    Suitable for: IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Chicken, Dog, Human, Xenopus laevis, Amphibian, Xenopus tropicalis
    Predicted to work with: Guinea pig
  • Immunogen

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

    DLKEEKDINNNVKKT

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control
    • WB: Recombinant human or mouse iNOS, cytokine stimulated RAW 264.7 cell lysate, cytokine stimulated rat fibroblast and LPS treated mouse spleen tissue lysates.

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
IHC-P Use at an assay dependent concentration. PubMed: 28186091
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

  • Function
    Produces 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 specificity
    Expressed in the liver, retina, bone cells and airway epithelial cells of the lung. Not expressed in the platelets.
  • Sequence similarities
    Belongs to the NOS family.
    Contains 1 FAD-binding FR-type domain.
    Contains 1 flavodoxin-like domain.
  • Information by UniProt
  • Database links
    see all
  • 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

Images

  • Western blot - Anti-iNOS antibody (ab3523)
    Western blot - Anti-iNOS antibody (ab3523)
    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
    All lanes : 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

  • Western blot - Anti-iNOS antibody (ab3523)
    Western blot - Anti-iNOS antibody (ab3523)This image is courtesy of an anonymous abreview.
    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
    All lanes : Goat Anti-rabbit IgG, HRP-linked at 1/2000 dilution

    Developed using the ECL technique.

    Exposure time: 5 minutes


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

    See Abreview

References

This product has been referenced in:
  • Zhou Z  et al. circRNA Mediates Silica-Induced Macrophage Activation Via HECTD1/ZC3H12A-Dependent Ubiquitination. Theranostics 8:575-592 (2018). Read more (PubMed: 29290828) »
  • Lankford KL  et al. Intravenously delivered mesenchymal stem cell-derived exosomes target M2-type macrophages in the injured spinal cord. PLoS One 13:e0190358 (2018). Read more (PubMed: 29293592) »

See all 89 Publications for this product

Publishing research using ab3523? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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53 reviews or Q&A

10
Votes: Highest First

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-iNOS antibody

 Poor
Abreviews
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
Read More
abreview image
Username

Abcam user community

Verified customer

Submitted May 21 2015

mouse liver samples
sees 3 bands between 120 and 220 kDa
which one is correct?
customer will send image for getting advice

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.

Western blot abreview for Anti-iNOS antibody

 Excellent
Abreviews
Application
Western blot
Sample
Mouse Tissue lysate - whole (Kidney)
Gel Running Conditions
Reduced Denaturing (4-12)
Loading amount
40 µg
Specification
Kidney
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
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Username

Dr. Wesley Konsavage

Verified customer

Submitted Dec 22 2016

Western blot abreview for Anti-iNOS antibody

 Excellent
Abreviews
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
Read More
abreview image
Username

Abcam user community

Verified customer

Submitted Mar 07 2016

Western blot abreview for Anti-iNOS antibody

 Good
Abreviews
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
Read More
abreview image
Username

Abcam user community

Verified customer

Submitted Mar 07 2016

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-iNOS antibody

 Good
Abreviews
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
Read More
abreview image
Username

Abcam user community

Verified customer

Submitted Nov 12 2015

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-iNOS antibody

 Good
Abreviews
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
Read More
abreview image
Username

Abcam user community

Verified customer

Submitted Oct 14 2015

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-iNOS antibody

 Good
Abreviews
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
Read More
abreview image
Username

Abcam user community

Verified customer

Submitted Nov 25 2014

Western blot abreview for Anti-iNOS antibody

 Excellent
Abreviews
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
Read More
abreview image
Username

Abcam user community

Verified customer

Submitted Jun 13 2014

Western blot abreview for Anti-iNOS antibody

 Excellent
Abreviews
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
Read More
abreview image
Username

Abcam user community

Verified customer

Submitted Jun 12 2014

1-10 of 53 Abreviews or Q&A

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