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    inos-antibody-ab3523.pdf

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Neuroscience Neurotransmission Nitric Oxide NOS
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Anti-iNOS antibody (ab3523)

  • Datasheet
  • SDS
Reviews (31)Q&A (33)References (199)

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Immunohistochemistry - Anti-iNOS antibody (ab3523)
  • Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody (ab3523)
  • Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody (ab3523)

Key features and details

  • Rabbit polyclonal to iNOS
  • Suitable for: ICC/IF, IHC
  • Reacts with: Mouse, Human
  • Isotype: IgG

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Protein
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Recombinant Human iNOS protein (ab135010)
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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 & Species

    Application Species
    ICC/IF
    Mouse
    Human
    IHC
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide corresponding to Mouse iNOS aa 1-100.
    Database link: P29477

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control

    • ICC: A549, NIH/3T3 cells. IHC-P: Human heart tissue.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 99% PBS
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurotransmission
    • Nitric Oxide
    • NOS
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Cancer
    • Cancer Metabolism
    • Cellular metabolic process
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Positive Controls

    • NIH/3T3 whole cell lysate (ab7179)
  • Recombinant Protein

    • Recombinant Human iNOS protein (ab135010)
  • Related Products

    • Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)

Applications

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Mouse
Human
IHC
Human
Application Abreviews Notes
ICC/IF (2)
1/50.
IHC
1/20.
Notes
ICC/IF
1/50.
IHC
1/20.

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.
  • Target information above from: UniProt accession P35228 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 4843 Human
    • Entrez Gene: 18126 Mouse
    • Omim: 163730 Human
    • SwissProt: P35228 Human
    • SwissProt: P29477 Mouse
    • Unigene: 709191 Human
    • Unigene: 2893 Mouse
    • 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

    • Immunohistochemistry - Anti-iNOS antibody (ab3523)
      Immunohistochemistry - Anti-iNOS antibody (ab3523)

      Immunohistochemistry analysis of human heart tissue stained for iNOS without (negative control) or using ab3523 at 1/200 dilution overnight at 4°C in a humidified chamber, followed biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor.

      To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature.

    • Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody (ab3523)
      Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody (ab3523)

      Immunofluorescence analysis of A549 (human lung carcinoma cell line) whole cells labelling iNOS (Left panel: green) without (control) or using ab3523 at 1/20 dilution overnight at 4°C, followed DyLight-488 conjugated secondary antibody. Counter stain: F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

      Cells were grown on chamber slides and fixed with formaldehyde prior to staining.

       

    • Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody (ab3523)
      Immunocytochemistry/ Immunofluorescence - Anti-iNOS antibody (ab3523)

      Immunofluorescence analysis of NIH/3T3 (mouse embryo fibroblast cell line) whole cells labelling iNOS (Left panel: green) without (control) or using ab3523 at 1/20 dilution overnight at 4°C, followed DyLight-488 conjugated secondary antibody. Counter stain: F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.

      Cells were grown on chamber slides and fixed with formaldehyde prior to staining.

       

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS
  • References (199)

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

    ab3523 has been referenced in 199 publications.

    • Ahmad N  et al. Imperatorin suppresses IL-1ß-induced iNOS expression via inhibiting ERK-MAPK/AP1 signaling in primary human OA chondrocytes. Int Immunopharmacol 85:106612 (2020). PubMed: 32450530
    • Miguel-Jiménez S  et al. Does Melatonin Exert Its Effect on Ram Sperm Capacitation Through Nitric Oxide Synthase Regulation? Int J Mol Sci 21:N/A (2020). PubMed: 32197481
    • Zhao JM  et al. Anserine and glucosamine supplementation attenuates the levels of inflammatory markers in rats with rheumatoid arthritis. AMB Express 10:57 (2020). PubMed: 32198574
    • Wang J  et al. Suberoylanilide hydroxamic acid alleviates orthotopic liver transplantation-induced hepatic ischemia-reperfusion injury by regulating the AKT/GSK3ß/NF-?B and AKT/mTOR pathways in rat Kupffer cells. Int J Mol Med 45:1875-1887 (2020). PubMed: 32236599
    • McKelvey KJ  et al. Temporal and spatial modulation of the tumor and systemic immune response in the murine Gl261 glioma model. PLoS One 15:e0226444 (2020). PubMed: 32240177
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 64 Abreviews or Q&A

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

    Poor
    Abreviews
    Abreviews
    abreview image
    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

    Abcam user community

    Verified customer

    Submitted May 21 2015

    Question

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

    Read More

    Abcam community

    Verified customer

    Asked on Feb 09 2012

    Answer

    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.

    Read More

    Abcam Scientific Support

    Answered on Feb 09 2012

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

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (kidney)
    Antigen retrieval step
    Enzymatic - Buffer/Enzyme Used: 0.05%trypsin
    Permeabilization
    No
    Specification
    kidney
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 18°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 25 2021

    Immunohistochemistry (Frozen sections) abreview for Anti-iNOS antibody

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Rat Tissue sections (Dorsal Root Ganglion)
    Permeabilization
    Yes - triton
    Specification
    Dorsal Root Ganglion
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
    Fixative
    Paraformaldehyde
    Read More

    Xiang Xu

    Verified customer

    Submitted Dec 27 2019

    Western blot abreview for Anti-iNOS antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (primary cardiomyocytes)
    Gel Running Conditions
    Reduced Denaturing (4-12 %)
    Loading amount
    20 µg
    Treatment
    LPS 1 µg/ml
    Specification
    primary cardiomyocytes
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Dr. Barbara Thaler

    Verified customer

    Submitted Jul 04 2019

    Western blot abreview for Anti-iNOS antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (HL-1 cardiomyocytes)
    Gel Running Conditions
    Reduced Denaturing (4-12 %)
    Loading amount
    40 µg
    Treatment
    LPS 1 µg/ml
    Specification
    HL-1 cardiomyocytes
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Dr. Barbara Thaler

    Verified customer

    Submitted Jul 03 2019

    Western blot abreview for Anti-iNOS antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Primary murine microglia)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    50 µg
    Treatment
    1µM LPA for different time points
    Specification
    Primary murine microglia
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More

    Dr. Lisha Joshi

    Verified customer

    Submitted Jun 11 2019

    Immunocytochemistry/ Immunofluorescence abreview for Anti-iNOS antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Primary murine microglia)
    Permeabilization
    Yes - Triton X
    Specification
    Primary murine microglia
    Blocking step
    BSA as blocking agent for 40 minute(s) · Concentration: 5% · Temperature: 4°C
    Fixative
    Paraformaldehyde
    Read More

    Dr. Lisha Joshi

    Verified customer

    Submitted Jun 11 2019

    Western blot abreview for Anti-iNOS antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (skin)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    10 µg
    Treatment
    UV B (100mJ/cm2)
    Specification
    skin
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 05 2019

    Immunohistochemistry (Frozen sections) abreview for Anti-iNOS antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Axolotl Tissue sections (Arm)
    Permeabilization
    No
    Specification
    Arm
    Blocking step
    DAKO serum free blocking as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 0µg/mL · Temperature: 25°C
    Fixative
    N/a
    Read More

    Miss. Crystal Jones

    Verified customer

    Submitted Nov 02 2018

    1-10 of 64 Abreviews or Q&A

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