Overview

  • Product name
    Anti-iNOS antibody [EPR16635]
    See all iNOS primary antibodies
  • Description
    Rabbit monoclonal [EPR16635] to iNOS
  • Host species
    Rabbit
  • Specificity
    This antibody shows low affinity on human samples.
  • Tested applications
    Suitable for: WB, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fragment within Human iNOS aa 1-200. The exact sequence is proprietary.
    Database link: P35228

  • Positive control
    • WB: RAW 264.7 treated with 0.1 µg/mL LPS for 6 hours, HepG2 treated with 10 µg/mL LPS for 6 hours,whole cell lysates; Human fetal brain lysate. ICC/IF: RAW 264.7 cells treated with LPS (0.1 µg/mL), for 6 hours. IP: RAW 264.7 whole cell lysate treated with 1µg/mL LPS for 24h.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab178945 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 1/1000. Detects a band of approximately 131 kDa (predicted molecular weight: 131 kDa).

To detect iNOS expression, macrophages need to be treated with LPS (0.1 - 1 ug/ml).

ICC/IF 1/500.

To detect iNOS expression, macrophages need to be treated with LPS (0.1 - 1 ug/ml).

IP 1/100.

To detect iNOS expression, macrophages need to be treated with LPS (0.1 - 1 ug/ml).

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
  • 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

  • All lanes : Anti-iNOS antibody [EPR16635] (ab178945) at 1/20000 dilution

    Lane 1 : Untreated RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 2 : RAW 264.7 whole cell lysate treated with 0.1 µg/mL LPS for 6 hours

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 131 kDa
    Observed band size: 131 kDa


    Exposure time: 30 seconds


    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/Immunofluorescence analysis of RAW 264.7 non-treated and LPS treated (0.1 µg/mL) cells labelling iNOS with ab178945 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. The cells were co-stained with ab195889, Alexa Fluor® 594 conjugated anti-alpha tubulin (1/200). Nuclei counterstained with DAPI (blue).

    Secondary antibody only controls performed on non-treated and treated cells.

  • iNOS was immunoprecipitated from 1mg of RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate treated with 1 µg/ml LPS for 24h with ab178945 at 1/100 dilution.

    Lane 1: RAW 264.7 whole cell lysate treated with 1 µg/ml LPS for 24h,10ug (Input).

    Lane 2: ab178945 IP in RAW 264.7 whole cell lysate treated with 1 µg/ml LPS for 24h.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab178945 in RAW 264.7 whole cell lysate treated with 1 µg/ml LPS for 24h.

    Western blot was performed from the immunoprecipitate using ab178945 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/1500.

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

    Exposure time: 30 seconds.

  • All lanes : Anti-iNOS antibody [EPR16635] (ab178945) at 1/500 dilution

    Lane 1 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates
    Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell) treated with 10ug/ml lipopolysaccharides for 6 hours whole cell lysates
    Lane 3 : THP-1 (Human monocytic leukemia monocyte) whole cell lysates
    Lane 4 : THP-1 (Human monocytic leukemia monocyte) treated with 100ng/ml lipopolysaccharides for 3 hours whole cell lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/200000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 131 kDa


    Exposure time: 3 minutes

References

This product has been referenced in:
  • Yuan C  et al. OAB-14, a bexarotene derivative, improves Alzheimer's disease-related pathologies and cognitive impairments by increasing ß-amyloid clearance in APP/PS1 mice. Biochim Biophys Acta Mol Basis Dis 1865:161-180 (2019). Read more (PubMed: 30389579) »
  • Lu Y  et al. Sertraline ameliorates inflammation in CUMS mice and inhibits TNF-a-induced inflammation in microglia cells. Int Immunopharmacol 67:119-128 (2019). Read more (PubMed: 30544065) »
See all 22 Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews

Application
Western blot
Sample
Human Tissue lysate - whole (skin)
Gel Running Conditions
Reduced Denaturing (10% Acrylamide gel)
Loading amount
20 µg
Treatment
UV B 100mJ/cm2
Specification
skin
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Mr. Jeong-Yong Park

Verified customer

Submitted Mar 15 2019

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Melanoma infiltrated tissue)
Permeabilization
No
Specification
Melanoma infiltrated tissue
Blocking step
(agent) for 10 minute(s) · Concentration: 0.25% · Temperature: 23°C
Fixative
Acetone

Abcam user community

Verified customer

Submitted Oct 25 2016

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