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    enos-antibody-epr19296-ab199956.pdf

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Neuroscience Neurotransmission Nitric Oxide NOS
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RecombinantRabMAb

Recombinant Anti-eNOS antibody [EPR19296] (ab199956)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (3) Submit a question References (26)

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Western blot - Anti-eNOS antibody [EPR19296] (ab199956)
  • Western blot - Anti-eNOS antibody [EPR19296] (ab199956)
  • Western blot - Anti-eNOS antibody [EPR19296] (ab199956)
  • Western blot - Anti-eNOS antibody [EPR19296] (ab199956)
  • Western blot - Anti-eNOS antibody [EPR19296] (ab199956)
  • Immunoprecipitation - Anti-eNOS antibody [EPR19296] (ab199956)
  • Anti-eNOS antibody [EPR19296] (ab199956)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19296] to eNOS
  • Suitable for: IP, WB
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

You may also be interested in

Protein
Product image
Recombinant Human eNOS protein (ab112329)
Primary
Product image
Anti-eNOS (phospho S1177) antibody [EPR20991] (ab215717)
Primary
Product image
Anti-eNOS (phospho S1177) antibody [EPR20991] - BSA and Azide free (ab230158)

View more associated products

Overview

  • Product name

    Anti-eNOS antibody [EPR19296]
    See all eNOS primary antibodies
  • Description

    Rabbit monoclonal [EPR19296] to eNOS
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human fetal heart, fetal kidney and fetal liver lysates; Mouse brain, heart, kidney and spleen lysates; Rat brain, heart and spleen lysates; whole cell lysate from HepG2 cells treated with 10 µg/ml LPS for 2 hours and 6 hours; DDDDK tagged human eNOS full length recombinant protein (aa2-1203) ab198066. IP: HepG2 whole cell lysate.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

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

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR19296
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurotransmission
    • Nitric Oxide
    • NOS
    • Cardiovascular
    • Blood
    • Platelets
    • Cell Biology
    • Other Antibodies
    • Oxidative Stress
    • Cardiovascular
    • Vasculature
    • Vasodilation
    • Nitric oxide associated
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia
    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Oxidative stress

Associated products

  • Alternative Versions

    • Anti-eNOS antibody [EPR19296] - BSA and Azide free (ab240352)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Recombinant Human eNOS protein (Tagged) (ab198066)
  • Recombinant Protein

    • Recombinant Human eNOS protein (ab112329)
  • Related Products

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab199956 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
IP (1)
1/50.
WB (2)
1/1000. Detects a band of approximately 140 kDa (predicted molecular weight: 133 kDa).
Notes
IP
1/50.
WB
1/1000. Detects a band of approximately 140 kDa (predicted molecular weight: 133 kDa).

Target

  • Function

    Produces nitric oxide (NO) which is implicated in vascular smooth muscle relaxation through a cGMP-mediated signal transduction pathway. NO mediates vascular endothelial growth factor (VEGF)-induced angiogenesis in coronary vessels and promotes blood clotting through the activation of platelets.
    Isoform eNOS13C: Lacks eNOS activity, dominant-negative form that may down-regulate eNOS activity by forming heterodimers with isoform 1.
  • Tissue specificity

    Platelets, placenta, liver and kidney.
  • Involvement in disease

    Variation in NOS3 seem to be associated with susceptibility to coronary spasm.
  • Sequence similarities

    Belongs to the NOS family.
    Contains 1 FAD-binding FR-type domain.
    Contains 1 flavodoxin-like domain.
  • Post-translational
    modifications

    Phosphorylation by AMPK at Ser-1177 in the presence of Ca(2+)-calmodulin (CaM) activates activity. In absence of Ca(2+)-calmodulin, AMPK also phosphorylates Thr-495, resulting in inhibition of activity (By similarity). Phosphorylation of Ser-114 by CDK5 reduces activity.
  • Cellular localization

    Cell membrane. Membrane, caveola. Cytoplasm, cytoskeleton. Golgi apparatus. Specifically associates with actin cytoskeleton in the G2 phase of the cell cycle and which is favored by interaction with NOSIP and results in a reduced enzymatic activity.
  • Target information above from: UniProt accession P29474 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 4846 Human
    • Entrez Gene: 18127 Mouse
    • Entrez Gene: 24600 Rat
    • Omim: 163729 Human
    • SwissProt: P29474 Human
    • SwissProt: P70313 Mouse
    • SwissProt: Q62600 Rat
    • Unigene: 647092 Human
    • Unigene: 258415 Mouse
    • Unigene: 44265 Rat
    see all
  • Alternative names

    • cNOS antibody
    • Constitutive NOS antibody
    • EC NOS antibody
    • EC-NOS antibody
    • ecNOS antibody
    • Endothelial nitric oxidase synthase antibody
    • Endothelial nitric oxide synthase antibody
    • Endothelial nitric oxide synthase 3 antibody
    • Endothelial NOS antibody
    • eNOS antibody
    • Nitric oxide synthase 3 (endothelial cell) antibody
    • Nitric oxide synthase 3 antibody
    • Nitric oxide synthase 3 endothelial cell antibody
    • Nitric oxide synthase endothelial antibody
    • Nitric oxide synthase, endothelial antibody
    • NOS 3 antibody
    • NOS III antibody
    • NOS type III antibody
    • NOS3 antibody
    • NOS3_HUMAN antibody
    • NOSIII antibody
    see all

Images

  • Western blot - Anti-eNOS antibody [EPR19296] (ab199956)
    Western blot - Anti-eNOS antibody [EPR19296] (ab199956)
    All lanes : Anti-eNOS antibody [EPR19296] (ab199956) at 1/1000 dilution

    Lane 1 : Human fetal heart lysate
    Lane 2 : Human fetal kidney lysate
    Lane 3 : Human fetal liver lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 133 kDa
    Observed band size: 140 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    eNOS is shown to be expressed in platelets, placenta, liver and kidney (UniProt entry P29474). It also plays an important role in the cardiovascular system. (PMID: 15210449 & 15936740).

  • Western blot - Anti-eNOS antibody [EPR19296] (ab199956)
    Western blot - Anti-eNOS antibody [EPR19296] (ab199956)
    All lanes : Anti-eNOS antibody [EPR19296] (ab199956) at 1/1000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Mouse kidney lysate
    Lane 4 : Mouse spleen lysate
    Lane 5 : Rat brain lysate
    Lane 6 : Rat heart lysate
    Lane 7 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 133 kDa
    Observed band size: 140 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1, 2, 3, 4, 6 and 7: 30 seconds; Lane 5: 2 minutes.

    eNOS is shown to be expressed in platelets, placenta, liver and kidney (UniProt entry P29474). It also plays an important role in the cardiovascular system. (PMID: 15210449 & 15936740).

  • Western blot - Anti-eNOS antibody [EPR19296] (ab199956)
    Western blot - Anti-eNOS antibody [EPR19296] (ab199956)
    All lanes : Anti-eNOS antibody [EPR19296] (ab199956) at 1/1000 dilution

    Lane 1 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
    Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 133 kDa
    Observed band size: 140 kDa why is the actual band size different from the predicted?



    Exposure Time: Lane1: 40 seconds; Lane2: 180 seconds

    The expression profile observed in HeLa is consistent with the literature (PMID: 19925457).
    Negative control: HeLa (PMID: 19925457)

  • Western blot - Anti-eNOS antibody [EPR19296] (ab199956)
    Western blot - Anti-eNOS antibody [EPR19296] (ab199956)
    All lanes : Anti-eNOS antibody [EPR19296] (ab199956) at 1/5000 dilution

    Lane 1 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
    Lane 2 : Whole cell lysate from HepG2 cells treated with 10 µg/ml LPS for 2 hours
    Lane 3 : Whole cell lysate from HepG2 cells treated with 10 µg/ml LPS for 6 hours

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 133 kDa
    Observed band size: 140 kDa why is the actual band size different from the predicted?


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression profile observed is consistent with what has been described in the literature (PMID:17855661 & 11004215).

  • Western blot - Anti-eNOS antibody [EPR19296] (ab199956)
    Western blot - Anti-eNOS antibody [EPR19296] (ab199956)
    All lanes : Anti-eNOS antibody [EPR19296] (ab199956) at 1/1000 dilution

    Lane 1 : Recombinant Human eNOS protein (Tagged) (ab198066) at 0.01 µg
    Lane 2 : Recombinant Human iNOS protein (ab135010) at 0.05 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 133 kDa
    Observed band size: 140 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 1 second; Lane 2: 3 minutes.

     

  • Immunoprecipitation - Anti-eNOS antibody [EPR19296] (ab199956)
    Immunoprecipitation - Anti-eNOS antibody [EPR19296] (ab199956)

    eNOS was immunoprecipitated from 1 mg of HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate with ab199956 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab199956 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HepG2 whole cell lysate 10 μg (Input).

    Lane 2: ab199956 IP in HepG2 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab199956 in HepG2 whole cell lysate.

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

    Exposure time: 3 seconds.

  • Anti-eNOS antibody [EPR19296] (ab199956)
    Anti-eNOS antibody [EPR19296] (ab199956)

Protocols

  • Western blot protocols
  • Immunoprecipitation protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (26)

Publishing research using ab199956? Please let us know so that we can cite the reference in this datasheet.

ab199956 has been referenced in 26 publications.

  • Liu Y  et al. Insulin Resistance and Pellino-1 Mediated Decrease in the Activities of Vasodilator Signaling Contributes to Sunitinib-Induced Hypertension. Front Pharmacol 12:617165 (2021). PubMed: 33841146
  • Xu S  et al. The role of RNA m6A methylation in the regulation of postnatal hypoxia-induced pulmonary hypertension. Respir Res 22:121 (2021). PubMed: 33902609
  • Shen B  et al. LncRNA GAS5 Silencing Attenuates Oxygen-Glucose Deprivation/Reperfusion-Induced Injury in Brain Microvascular Endothelial Cells via miR-34b-3p-Dependent Regulation of EPHA4. Neuropsychiatr Dis Treat 17:1667-1678 (2021). PubMed: 34079264
  • Maldonado M  et al. The consequences of a high-calorie diet background before calorie restriction on skeletal muscles in a mouse model. Aging (Albany NY) 13:16834-16858 (2021). PubMed: 34166224
  • Yu J  et al. Roxadustat prevents Ang II hypertension by targeting angiotensin receptors and eNOS. JCI Insight 6:N/A (2021). PubMed: 34403364
View all Publications for this product

Customer reviews and Q&As

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1-3 of 3 Abreviews

Western blot abreview for Anti-eNOS antibody [EPR19296]

Average
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Rat Tissue lysate - whole (cardiac muscle)
Gel Running Conditions
Non-reduced Denaturing
Loading amount
20 µg
Specification
cardiac muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 24 2021

Western blot abreview for Anti-eNOS antibody [EPR19296]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Human Umbilical Vein Endothelial Cells)
Gel Running Conditions
Non-reduced Denaturing
Loading amount
20 µg
Specification
Human Umbilical Vein Endothelial Cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 09 2021

Immunoprecipitation abreview for Anti-eNOS antibody [EPR19296]

Good
Abreviews
Abreviews
abreview image
Application
Immunoprecipitation
Sample
Mouse Tissue lysate - whole (liver)
Total protein in input
2000 µg
Immuno-precipitation step
Other - Protein G dynabeads
Specification
liver
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 10 2018

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