Purified anti-NOS3 (eNOS) Antibody

Pricing & Availability
Clone
W22110B (See other available formats)
Regulatory Status
RUO
Other Names
Nitric oxide synthase 3 (NOS3), Endothelial Nos (eNos), constitutive NOS (cNOS)
Isotype
Rat IgG2b, κ
Ave. Rating
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Product Citations
publications
A.
W22110B_PURE_NOS3_-IHC-P-_112023
IHC staining of Purified anti-NOS3 (eNOS) (clone W22110B) on formalin-fixed paraffin-embedded human placenta tissue. Following antigen retrieval using Sodium Citrate H.I.E.R., 1X (Cat. No. 928502), the tissue was incubated without (panel A) and with (panel B) 5 µg/mL of Purified anti-NOS3 (eNOS) (clone W22110B) followed by incubation with Alexa Fluor® 647 Goat anti-rat IgG (Cat. No. 405416) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801). Images were captured with a 40X objective. Scale bar: 50 µm
  • A.
W22110B_PURE_NOS3_-IHC-P-_112023
    IHC staining of Purified anti-NOS3 (eNOS) (clone W22110B) on formalin-fixed paraffin-embedded human placenta tissue. Following antigen retrieval using Sodium Citrate H.I.E.R., 1X (Cat. No. 928502), the tissue was incubated without (panel A) and with (panel B) 5 µg/mL of Purified anti-NOS3 (eNOS) (clone W22110B) followed by incubation with Alexa Fluor® 647 Goat anti-rat IgG (Cat. No. 405416) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801). Images were captured with a 40X objective. Scale bar: 50 µm
  • B.
W22110B_PURE_NOS3_-ICC_112023
    EA.hy926 cells (positive cell line) were fixed with 4% PFA Fixation Buffer (Cat. No. 420801) for 10 minutes and permeabilized with 0.5% Triton X-100 for 10 minutes, and blocked with 5% FBS for 1 hour at room temperature. Cells were then stained with (panel B) or without (panel A) 5 µg/mL of Purified anti-NOS3 (eNOS) (clone W22110B), followed by incubation with Alexa Fluor® 594 Goat anti-rat IgG (Cat. No. 405422) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801). The images were captured on a Revvity Operetta CLS™ High Content Analysis System with a 60X objective. Scale bar: 50 µm
  • C.
W22110B_PURE_NOS3_-WB_112023
    Whole cell extracts (15 µg total protein) from indicated cell lines and tissue were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 0.25 µg/mL of Purified anti-NOS3 (eNOS) (clone W22110B) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH (Cat. No. 607904) was used as a loading control at a 1:50000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • D.W22110B_PURE_NOS3_Antibody_ICFC_4_032724
    EA.hy926 cells (positive control, filled histogram) or HeLa cells (negative control, open histogram) were fixed and permeabilized using Cyto-Fast™ Fix/Perm Buffer Set (Cat. No. 426803) and stained with Purified anti-NOS3 (eNOS) (clone W22110B) or Purified Rat IgG2b, κ isotype control (open histogram, dashed line) (representative histogram for both cell lines) (Cat. No. 400601), followed by Alexa Fluor® 647 Goat anti-rat IgG (Cat. No. 405416).
Cat # Size Price Save
626751 25 µg ¥38,500
626752 100 µg ¥100,100
Description

Nitric oxide synthase 3 (NOS3), also commonly known as endothelial nitric oxide synthase (eNOS), is a critical enzyme primarily found in the endothelial cells lining the blood vessels. Its main function is to catalyze the formation of nitric oxide (NO) from L-arginine. NO is a signaling molecule with profound effects on vascular health and overall physiology, regulating blood vessel tone and diameter in response to various physiological cues including shear stress, neurotransmitters, and hormones. NO relaxes and dilates blood vessels, thereby promoting proper blood flow and reducing blood pressure. Additionally, NOS3-derived NO helps prevent the formation of blood clots, reduces inflammation in the blood vessel walls, and maintains vascular homeostasis, contributing significantly to cardiovascular health. Dysfunction or impaired activity of NOS3 is implicated in various cardiovascular diseases, making it an important target for therapeutic interventions aimed at preserving vascular function.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Recombinant KLH-conjugated peptide fragment of human NOS3
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

IHC-P - Quality tested
ICC, WB, ICFC - Verified

Recommended Usage

Each lot of this antibody is quality control tested by formalin-fixed paraffin-embedded immunohistochemical staining. For immunohistochemistry, a concentration range of 1.0 - 10.0 µg/mL is suggested. For immunocytochemistry, a concentration range of 1.25 - 10.0 μg/mL is recommended. For western blotting, the suggested use of this reagent is 0.25 - 1.0 µg/mL. For intracellular flow cytometric staining, the suggested use of this reagent is ≤ 0.5 µg per million cells in 100 µL volume. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This antibody clone, W22110B, is not suitable for ICFC (Intracellular Staining for Flow Cytometry).

This antibody clone, W22110B, does not cross-react with mouse NOS3 in Western Blot application.

For immunohistochemistry (IHC-P), either Citrate Buffer, 10X (Cat. No. 420902) or Tris-EDTA pH 9.0 is recommended for antigen retrieval system.

For immunocytochemistry (ICC), Fixation Buffer (Cat. No. 420801) or 4% paraformaldehyde fixation followed by permeabilization with 0.5% Triton X-100 is recommended. Fixation/permeabilization with methanol is not recommended.

Antigen Details

Structure
NOS3 is a 1,205 amino acid protein with a predicted molecular weight of 133.3 kD.
Distribution

Cytoplasmic, vascular endothelial cells

Function
Enzyme, nitric oxide (NO) synthesis from L-arginine. Transcriptionally regulated by SP1, SP3, ETS-1, YY1.
Interaction
NOS3 is regulated via binding of its cofactor BH4, which is required to efficiently produce NO
Cell Type
Endothelial cells
Biology Area
Cardiovascular Biology
Molecular Family
Enzymes and Regulators
Antigen References
  1. Mineo C and Shaul PW. 2012. Adv Exp Med Biol. 729:51-62. 
  2. Hong FF, et al. 2019. Inflamm Res. 68:429-441. 
  3. Siragusa M and Fleming I. 2016. Pfulgers Arch. 468:1125-1137. 
  4. Musicki B and Burnett AL 2006. Exp Bil Med. 231:154-65.
Gene ID
4846 View all products for this Gene ID
UniProt
View information about NOS3 on UniProt.org

Related FAQs

There are no FAQs for this product.
Go To Top Version: 2    Revision Date: 03/27/2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

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Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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