製品の概要

  • 製品名Anti-iNOS antibody
    iNOS 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to iNOS
  • 特異性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.
  • アプリケーション適用あり: IHC-Fr, Electron Microscopy, WB, Flow Cyt, IHC-P, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Rat
  • 免疫原

    Synthetic peptide:

    CFSYGAKKGSALEEPKATRL

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

  • ポジティブ・コントロール
    • Lung; Raw 264.7 whole cell lysate + PMA + LPS + Brefeldin A

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
IHC-Fr Use at an assay dependent concentration. PubMed: 18523253
Electron Microscopy Use at an assay dependent concentration. PubMed: 21071693
WB Use a concentration of 5 µg/ml. Detects a band of approximately 140 kDa (predicted molecular weight: 131 kDa).
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

ターゲット情報

  • 機能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.
  • 組織特異性Expressed in the liver, retina, bone cells and airway epithelial cells of the lung. Not expressed in the platelets.
  • 配列類似性Belongs to the NOS family.
    Contains 1 FAD-binding FR-type domain.
    Contains 1 flavodoxin-like domain.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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

Anti-iNOS antibody 画像

  • 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) 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

  • 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.
  • 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

  • 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

  • ab15323 staining iNOS in Mouse lung by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 1% BSA for 30 minutes at 27°C. Samples were incubated with primary antibody (1/200) for 1 hour at 27°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit polyclonal was used as the secondary antibody (1/500). Heat antigen retrieval (in Citrate buffer) was necessary.

    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.

Anti-iNOS antibody (ab15323) 使用論文

This product has been referenced in:
  • Misiorek JO  et al. Keratin 8-deletion induced colitis predisposes to murine colorectal cancer enforced by the inflammasome and IL-22 pathway. Carcinogenesis 37:777-86 (2016). Read more (PubMed: 27234655) »
  • Zhu W  et al. Glioma-mediated microglial activation promotes glioma proliferation and migration: roles of Na+/H+ exchanger isoform 1. Carcinogenesis 37:839-51 (2016). Read more (PubMed: 27287871) »

See all 48 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry free floating
Sample Mouse Tissue sections (brain)
Specification brain
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投稿 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
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投稿 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
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投稿 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
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投稿 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
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投稿 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
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投稿 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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Vielen Dank für Ihren Anruf und für Ihr Interesse an unseren Produkten.
 
Ich habe die Homologie der Immunogene unsere Anti iNOS Antikörper mit Schwein verglichen und habe folgende mögliche Antikörper für Sie:
ab129372 : 85%
ab13691...

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Thank you for contacting us and your interest in our products. As discussed over the phone, I have checked the concentration of the antibodies and lot numbers you provided. The concentrations are as follows:

ab52897, GR5286-6, 0.165 mg/ml;
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"