概述

  • 产品名称
  • 描述
    兔多克隆抗体to iNOS
  • 特异性
    We have internal testing data to indicate that this antibody may not react with human and therefore we cannot guarantee it for use in this species. The immunogen for this product has 84% homology with the Rat iNOS protein.
  • 经测试应用
    适用于: IHC-Fr, Electron Microscopy, WB, Flow Cyt, IHC-P, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat
  • 免疫原

    Synthetic peptide:

    CFSYGAKKGSALEEPKATRL

    , corresponding to C terminal amino acids 1125-1144 of Mouse iNOS.

  • 阳性对照
    • Lung; Raw 264.7 whole cell lysate + PMA + LPS + Brefeldin A

性能

应用

Our Abpromise guarantee covers the use of ab15323 in the following tested applications.

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

应用 Ab评论 说明
IHC-Fr Use at an assay dependent concentration. PubMed: 18523253
Electron Microscopy Use at an assay dependent concentration. PubMed: 21071693
WB Use at an assay dependent concentration. Detects a band of approximately 140 kDa (predicted molecular weight: 131 kDa).

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.

Flow Cyt Use at an assay dependent concentration.
IHC-P 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration. PubMed: 20584290

靶标

  • 功能
    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.
  • 组织特异性
    Expressed in the liver, retina, bone cells and airway epithelial cells of the lung. Not expressed in the platelets.
  • 序列相似性
    Belongs to the NOS family.
    Contains 1 FAD-binding FR-type domain.
    Contains 1 flavodoxin-like domain.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

图片

  • All lanes : Anti-iNOS antibody (ab15323) at 1 µg/ml

    Lane 1 : Raw264.7
    Lane 2 : Raw264.7 + LPS (1 ug per mL; 18 hours)

    Lysates/proteins at 20 µg per lane.

    Secondary
    IR- goat anti-rabbit (green) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 131 kDa

    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 (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.

  • ab15323 staining macrophages in mouse spleen sections by IHC-P. The tissue was fixed with formaldehyde and a heat mediated antigen retrival step was performed with citric acid pH 6. Blocking of the sample was done with 1%BSA for 10 minutes at 21°C, followed by staining with ab15323 at 1/50 in TBS/BSA/azide for 16h at 21°C. A biotinylated goat anti-rabbit polyclonal antibody at 1/200 was used as the secondary antibody.

    See Abreview

  • ab15323 staining iNOS in Mouse NIH3T3 cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton and blocked with 10% serum for 30 minutes at 24°C. Samples were incubated with primary antibody (1/100) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated Donkey anti-rabbit polyclonal was used as the secondary antibody (1/500).

    See Abreview

  • ab15323 staining iNOS in Mouse whole blood cells by Flow Cytometry. The sample was incubated with the undiluted primary antibody for 30 minutes at 25°C. ab97050, FITC-conjugated Goat anti-rabbit IgG (undiluted) was used as the secondary antibody.

    See Abreview

  • We performed immunostaining for the three NOS isoforms to examine their localization in mesenteric arteries of WT and the three genotypes of NOS-/- mice. In WT mice, the immunoreactivity of eNOS and nNOS, but not that of iNOS, was noted mainly in the endothelium, and in eNOS-/- and n/eNOS-/- mice, the expression of nNOS and iNOS was noted, respectively. In contrast, in n/i/eNOS-/- mice, none of the NOS isoforms were noted, as expected. Furthermore, we performed Western blot analysis to quantify the protein expression of NOS isoforms in the whole mesentery. In WT mice, the expression of both eNOS and nNOS was noted, whereas that of iNOS was minimal. In eNOS-/- mice, the expression of nNOS was again noted, and in n/eNOS-/- mice, the expression of iNOS was significantly up-regulated compared with WT or eNOS-/- mice. In n/i/eNOS-/- mice, none of the NOS isoforms were noted, as expected.
  • All lanes : Anti-iNOS antibody (ab15323) at 5 µg/ml

    Lane 1 : Raw 264.7 whole cell lysate + PMA
    Lane 2 : Raw 264.7 whole cell lysate + PMA + LPS + Brefeldin A

    Lysates/proteins at 20 µg per lane.

    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 131 kDa
    Additional bands at : 140 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 8 minutes
  • ab15323 staining rat liver (lower image) and spleen (upper image) sections by IHC-P. The tissue was fixed with formaldehyde and a heat mediated antigen retrival step was performed with citric acid pH 6. Blocking of the sample was done with 1%BSA for 10 minutes at 21°C, followed by staining with ab15323 at 1/100 in TBS/BSA/azide for 16h at 21°C. A biotinylated goat anti-rabbit polyclonal antibody at 1/200 was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-iNOS antibody (ab15323) at 5 µg/ml

    Lane 1 : Human recombinant iNOS protein
    Lane 2 : Mouse recombinant iNOS protein

    Lysates/proteins at 0.1 µg per lane.

    Secondary
    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


    Exposure time : 1 minute

    Membrane was blocked in 2% milk for 1 hour at RT. Primary anitbody was incubated at 4°C overnight.

文献

This product has been referenced in:
  • Wang Y  et al. Niaspan inhibits diabetic retinopathy-induced vascular inflammation by downregulating the tumor necrosis factor-a pathway. Mol Med Rep 15:1263-1271 (2017). Read more (PubMed: 28138697) »
  • Murray GM  et al. The bovine paranasal sinuses: Bacterial flora, epithelial expression of nitric oxide and potential role in the in-herd persistence of respiratory disease pathogens. PLoS One 12:e0173845 (2017). Read more (PubMed: 28282443) »

See all 101 Publications for this product

客户评价及客户问答

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (spleen)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9
Permeabilization
No
Specification
spleen
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 23°C
Fixative
10% NBF
Username

Abcam user community

Verified customer

提交于 Feb 27 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (spleen)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9
Permeabilization
No
Specification
spleen
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 23°C
Fixative
10% NBF
Username

Abcam user community

Verified customer

提交于 Feb 27 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (brain)
Specification
brain
Username

Abcam user community

Verified customer

提交于 Jun 28 2016

Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (Brain)
Permeabilization
No
Specification
Brain
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Apr 18 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse hepatocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Treatment
100 U/ml IL-1 beta
Specification
mouse hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

提交于 Mar 11 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
Sample
Human Cell (HUVEC)
Specification
HUVEC
Permeabilization
Yes - 0.1% Triton X100
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Sep 17 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Fixation
Formaldehyde
Permeabilization
Yes - 0.1% saponin
Sample
Mouse Cell (Mouse whole blood)
Specification
Mouse whole blood
Gating Strategy
CD14+ monocytes
Preparation
Cell harvesting/tissue preparation method: Whole blood collected in ACD
Sample buffer: None
Username

Abcam user community

Verified customer

提交于 Jul 17 2013

I have found that we can expect iNOS expression throughout the cell. In Saini.R. et.al. J Leukoc Biol. 2006 Mar;79(3):519-28. Epub 2005 Dec 30. PubMed PMID: 16387842. They show "iNOS and nNOS showed plasmalemmal, cytoplasmic, as well as nuclear d...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (NIH3T3)
Specification
NIH3T3
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
Username

Abcam user community

Verified customer

提交于 Apr 05 2013

Thank you for your response.

All our customer feedback, including complaints are monitored weekly by our in house technical support team. If a product is at fault the technical support team will consider removing the product from our catalogu...

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