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Anti-eNOS antibody [EPR26335-79] (ab300071) is a rabbit monoclonal antibody that is used to detect eNOS in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IHC-P, ICC/IF. Suitable for Mouse, Rat samples.



- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency

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Images

Western blot - Anti-eNOS antibody [EPR26335-79] (AB300071), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR26335-79] (AB300071), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR26335-79] (AB300071), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-eNOS antibody [EPR26335-79] (AB300071), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-eNOS antibody [EPR26335-79] (AB300071), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Abcam Recommends

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
Flow Cyt (Intra)IPWBICC/IFIHC-P
Human
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Mouse
Tested
Not recommended
Tested
Tested
Not recommended
Rat
Expected
Not recommended
Expected
Expected
Tested

Tested
Tested

Species
Mouse
Dilution info
1/500
Notes

-

Expected
Expected

Species
Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Mouse, Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/1000
Notes

-

Expected
Expected

Species
Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/50
Notes

-

Expected
Expected

Species
Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Rat
Dilution info
1/50
Notes

This antibody is not suitable for human species in IHC-P application.

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species
Mouse
Dilution info
-
Notes

This antibody is not suitable for human species in IHC-P application.

Species
Human
Dilution info
-
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

Select an associated product type

1 product for Alternative Product

Target data

Function

Produces nitric oxide (NO) which is implicated in vascular smooth muscle relaxation through a cGMP-mediated signal transduction pathway (PubMed:1378832). 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.

Alternative names

Nitric oxide synthase 3, Constitutive NOS, EC-NOS, NOS type III, cNOS, NOSIII, Endothelial NOS, eNOS, NOS3

Recommended products

Anti-eNOS antibody [EPR26335-79] (ab300071) is a rabbit monoclonal antibody that is used to detect eNOS in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IHC-P, ICC/IF. Suitable for Mouse, Rat samples.



- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR26335-79
Purification technique
Affinity purification Protein A
Specificity

Unsuitable for human, we recommend ab252439 and ab199956 for human. IHC-P unsuitable for mouse.

Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

What is this antibody validated in?


Anti-eNOS antibody [EPR26335-79] (ab300071) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF), in Mouse, Rat samples.

What is the molecular weight of eNOS?


Anti-eNOS [EPR26335-79] (ab300071) specifically detects a band for eNOS (UniProt: P70313) at a molecular weight of 132kDa.

Recommended positive controls


WB: Whole cell fresh lysates: bEnd.3 (mouse brain endothelioma). IHC-P: Rat spleen, colon, and stomach tissue.ICC/IF: bEnd.3 (mouse brain endothelioma endothelialpolyoma middle T antigen transformed). Flow cyt. intr.: bEnd.3 (mouse brain endothelioma endothelialpolyoma middle T antigen transformed).

Trial sizes available!


Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.Other related products


We have a range of other formats of antibody clone [EPR26335-79] also available for your convenience:
ab300071, Carrier free - Anti-eNOS antibody [EPR26335-79] (BSA and Azide free) ab300072



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

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

ENOS also known as endothelial nitric oxide synthase is an enzyme important for the production of nitric oxide (NO) in blood vessels. This protein with a molecular weight of approximately 133 kDa is expressed mostly in endothelial cells. eNOS plays a mechanical role in synthesizing NO from L-arginine a process requiring cofactors such as NADPH and oxygen. The activity of eNOS can be investigated through techniques such as Western blot with specific assays like phospho-eNOS ELISA available to measure its phosphorylated forms indicating activated states of the enzyme.

Biological function summary

ENOS contributes significantly to the regulation of vascular tone and blood flow by promoting vasodilation. It does not function alone; instead it forms complexes with other proteins to exert its full effect. eNOS activity influences the process of angiogenesis inflammation modulation and platelet aggregation. Through its ability to produce nitric oxide eNOS acts as a signaling molecule helping maintain vascular homeostasis.

Pathways

ENOS is a critical player in the NO signaling pathway and interacts intricately with the PI3K/Akt pathway. Nitric oxide produced by eNOS has a role in signaling cascades that lead to vascular dilation and reduced blood pressure. The PI3K/Akt pathway regulates eNOS activity via phosphorylation enhancing NO production. Other proteins like caveolin-1 and calmodulin modulate eNOS impacting these pathways' outcomes.

Associated diseases and disorders

ENOS is associated with cardiovascular conditions like atherosclerosis and hypertension. Dysfunction of eNOS can lead to reduced NO production impairing vasodilation and contributing to these diseases. In cardiovascular disorders eNOS interacts with proteins such as the angiotensin II type 1 receptor which can negatively impact its function exacerbating disease states. Investigating eNOS and its related proteins provides insight into potential therapeutic targets for improving cardiovascular health.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

5 product images

  • Western blot - Anti-eNOS antibody [EPR26335-79] (ab300071), expandable thumbnail

    Western blot - Anti-eNOS antibody [EPR26335-79] (ab300071)

    5% NFDM/TBST was used as blocking and diluting buffer.

    Exposure time:

    Lane 1: 10 seconds
    Lane 2: 3 minutes

    Negative control:RAW264.7 (PMID: 18607531)
    Lysates were freshly made and used in WB test immediately to minimize protein degradation.

    All lanes: Western blot - Anti-eNOS antibody [EPR26335-79] (ab300071) at 1/1000 dilution

    Lane 1: bEnd.3 (mouse brain endothelioma) whole cell fresh lysate at 20 µg

    Lane 2: RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell fresh lysate at 20 µg

    Secondary

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

    Predicted band size: 132 kDa, 133 kDa

    Observed band size: 140 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR26335-79] (ab300071), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR26335-79] (ab300071)

    Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling eNOS with ab300071 at 1/50 dilution (10.96 μg/ml), followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit. Positive staining on vascular endothelial cells of rat spleen was observed. The section was incubated with ab300071 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (BOND™ Polymer Refine Detection) was used.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR26335-79] (ab300071), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR26335-79] (ab300071)

    Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling eNOS with ab300071 at 1/50 dilution (10.96 μg/ml), followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit. Positive staining on vascular endothelial cells of rat stomach was observed. The section was incubated with ab300071 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

  • Immunocytochemistry/ Immunofluorescence - Anti-eNOS antibody [EPR26335-79] (ab300071), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-eNOS antibody [EPR26335-79] (ab300071)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized bEnd.3 (mouse brain endothelioma endothelialpolyoma middle T antigen transformed) cells lebeling eNOS with ab300071 at 1/50 dilution (10.96 μg/ml), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 μg/mL) (Green). Confocal image showing cytoplasm staining in bEnd.3 cell line. Negative control: RAW 264.7 (PMID: 18607531).

    Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5μg/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 μg/mL) dilution.

  • Flow Cytometry (Intracellular) - Anti-eNOS antibody [EPR26335-79] (ab300071), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-eNOS antibody [EPR26335-79] (ab300071)

    Flow cyt (Intra) analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage, Left) / bEnd.3 (Mouse brain endothelioma, Right) cells labeling eNOS with ab300071 at 1/500 dilution (0.1 μg) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control: RAW 264.7 (PMID: 18607531).

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Product protocols

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