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Rabbit Recombinant Monoclonal eNOS antibody. Suitable for mIHC, ICC/IF, IP, WB, Flow Cyt (Intra), IHC-P and reacts with Human samples.

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Images

Immunoprecipitation - Anti-eNOS antibody [EPR23750-3] (AB252439), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] (AB252439), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-eNOS antibody [EPR23750-3] (AB252439), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-eNOS antibody [EPR23750-3] (AB252439), expandable thumbnail
  • Western blot - Anti-eNOS antibody [EPR23750-3] (AB252439), expandable thumbnail

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
mIHCICC/IFIPWBFlow Cyt (Intra)IHC-P
Human
Tested
Tested
Tested
Tested
Tested
Tested
Mouse
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Rat
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended

Tested
Tested

Species

Human

Dilution info

1/200 - 1/1000

Notes

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

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/100

Notes

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

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/30

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/600

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/500

Notes

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

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

Species

Rat

Dilution info

-

Notes

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

Target data

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 eNOS13CLacks eNOS activity, dominant-negative form that may down-regulate eNOS activity by forming heterodimers with isoform 1.

Alternative names

Constitutive NOS, EC-NOS, Endothelial NOS, NOS type III, cNOS, eNOS, NOSIII, NOS3

Recommended products

Rabbit Recombinant Monoclonal eNOS antibody. Suitable for mIHC, ICC/IF, IP, WB, Flow Cyt (Intra), IHC-P and reacts with Human samples.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

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

EPR23750-3

Purification technique

Affinity purification Protein A

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

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

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.

11 product images

  • Immunoprecipitation - Anti-eNOS antibody [EPR23750-3] (ab252439), expandable thumbnail

    Immunoprecipitation - Anti-eNOS antibody [EPR23750-3] (ab252439)

    eNOS was immunoprecipitated from 0.35 mg EA.hy926 (human somatic cell hybrid endothelial) whole cell lysate with ab252439 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab252439 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

    Lane 1: EA.hy926 (human somatic cell hybrid endothelial) whole cell lysate 10 ug

    Lane 2: ab252439 IP in EA.hy926 (human somatic cell hybrid endothelial) whole cell lysate

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab252439 in EA.hy926 (human somatic cell hybrid endothelial) whole cell lysate

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

    Exposure time: 6 seconds.

    IP lysates were unboiled.

    All lanes: Immunoprecipitation - Anti-eNOS antibody [EPR23750-3] (ab252439)

    Predicted band size: 133 kDa

    Observed band size: 140 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] (ab252439), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] (ab252439)

    Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling eNOS with ab252439 at 1/500 (1.198 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Cytoplasmic staining on endothelial cells in human spleen. The section was incubated with ab252439 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 Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunocytochemistry/ Immunofluorescence - Anti-eNOS antibody [EPR23750-3] (ab252439), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-eNOS antibody [EPR23750-3] (ab252439)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HUVEC cells labelling eNOS with ab252439 at 1/100 (5.99 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HUVEC cells. Negative control: HeLa (PMID: 19559671). 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 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Flow Cytometry (Intracellular) - Anti-eNOS antibody [EPR23750-3] (ab252439), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-eNOS antibody [EPR23750-3] (ab252439)

    Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human cervix adenocarcinoma epithelial cell, left) /HUVEC (human umbilical vein endothelial cell, right) cells labelling eNOS with ab252439 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) isotype control (Black) 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) ab150077) at 1/2000 dilution was used as the secondary antibody.

    Negative control: Hela (PMID: 19559671).

  • Western blot - Anti-eNOS antibody [EPR23750-3] (ab252439), expandable thumbnail

    Western blot - Anti-eNOS antibody [EPR23750-3] (ab252439)

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

    HUVEC whole cell lysate in lane 1 is made freshly and used in WB test immediately to minimize protein degradation.

    Exposure time: 6 seconds.

    All lanes: Western blot - Anti-eNOS antibody [EPR23750-3] (ab252439) at 1/1000 dilution

    All lanes: HUVEC (human umbilical vein endothelial cell) whole cell lysate at 20 µg

    Secondary

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

    Predicted band size: 133 kDa

    Observed band size: 140 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] (ab252439), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] (ab252439)

    Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue labeling eNOS with abab252439 at 1/500 (1.198 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Cytoplasmic staining on endothelial cells in human lung carcinoma. The section was incubated with ab252439 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 Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Western blot - Anti-eNOS antibody [EPR23750-3] (ab252439), expandable thumbnail

    Western blot - Anti-eNOS antibody [EPR23750-3] (ab252439)

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

    Negative control: HeLa (PMID:19559671).

    Lysates were made freshly and used in WB test immediately to minimize protein degradation. Lysates were unboiled.

    Exposure time: 6 seconds.

    All lanes: Western blot - Anti-eNOS antibody [EPR23750-3] (ab252439) at 1/1000 dilution

    Lane 1: EA.hy926 (human somatic cell hybrid endothelial) whole cell lysate at 20 µg

    Lane 2: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Secondary

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

    Predicted band size: 133 kDa

    Observed band size: 140 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] (ab252439), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] (ab252439)

    Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling eNOS with ab252439 at 1/500 (1.198 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Cytoplasmic staining in human placenta. The section was incubated with ab252439 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 Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] (ab252439), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eNOS antibody [EPR23750-3] (ab252439)

    Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling eNOS with ab252439 at 1/500 (1.198 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Cytoplasmic staining on endothelial cells in human kidney. The section was incubated with ab252439 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 Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Multiplex immunohistochemistry - Anti-eNOS antibody [EPR23750-3] (ab252439), expandable thumbnail

    Multiplex immunohistochemistry - Anti-eNOS antibody [EPR23750-3] (ab252439)

    Fluorescence multiplex immunohistochemical analysis of human liver (formalin-fixed paraffin-embedded section). Panel A shows merged staining of anti-eNOS stained on endothelial cells (ab252439; red; Opal™570) at 1:1000 ( 1.004 μg/ml) [Panel B], anti-CD163 stained on Kupffer cells (Anti-CD163 antibody [EPR19518] - BSA and Azide free ab213612; green; Opal™520) at 1:8000 ( 0.13 μg/ml) [Panel B], and anti-Glucose Transporter GLUT2 stained on membrane of hepatocytes (Anti-Glucose Transporter GLUT2 antibody [EPR22946-74] ab234440; gray; Opal™690) at 1:200 ( 3.005 μg/ml) [Panel D] on human liver. DAPI was used as a nuclear counter stain. Followed by Opal Polymer HRP Ms + Rb secondary. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. The section was incubated in three rounds of staining: in the order of Anti-Glucose Transporter GLUT2 antibody [EPR22946-74] ab234440, Anti-CD163 antibody [EPR19518] - BSA and Azide free ab213612, and ab252439 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins was used.

  • Multiplex immunohistochemistry - Anti-eNOS antibody [EPR23750-3] (ab252439), expandable thumbnail

    Multiplex immunohistochemistry - Anti-eNOS antibody [EPR23750-3] (ab252439)

    Fluorescence multiplex immunohistochemical analysis of human liver (formalin-fixed paraffin-embedded section). Panel A shows merged staining of anti-MASP2 stained on cytoplasm of hepatocytes (Anti-MASP2 antibody [EPR23588-44] ab277520; gray; Opal™690) at 1:100 ( 5.22 μg/ml) [Panel B] , anti-CD163 stained on Kupffer cells (Anti-CD163 antibody [EPR19518] - BSA and Azide free ab213612; green; Opal™520) at 1:8000 ( 0.13 μg/ml) [Panel C], and anti-eNOS tained on endothelial cells (ab252439; red; Opal™570) at 1:200 ( 3.005 μg/ml) [Panel D] on human liver. DAPI was used as a nuclear counter stain. Followed by Opal Polymer HRP Ms + Rb secondary. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. The section was incubated in three rounds of staining: in the order of Anti-MASP2 antibody [EPR23588-44] ab277520, Anti-CD163 antibody [EPR19518] - BSA and Azide free ab213612, and ab252439 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins was used.

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