Anti-eNOS/NOS3 Antibody Picoband™

Product Name

Anti-eNOS/NOS3 Antibody Picoband™

View all eNOS Antibodies

SKU/Catalog Number

A01604-3

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-eNOS/NOS3 Antibody Picoband™ catalog # A01604-3. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human.

Storage & Handling

At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

Cite This Product

Anti-eNOS/NOS3 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01604-3)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human eNOS/NOS3 recombinant protein (Position: P34-Q1153).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A01604-3 is reactive to NOS3 in Human

Applications

A01604-3 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

140 kDa

Calculated molecular weight

133.275kDa

Background of eNOS

NOS3(Nitric Oxide Synthase 3), also called ENOS, anitric oxide synthasethat generatesNOinblood vesselsand is involved with regulatingvascular toneby inhibitingsmooth musclecontraction and plateletaggregation. The NOS3 gene is mapped on 7q36.1. Variations in this gene are associated with susceptibility tocoronary spasm. Fulton et al.(1999) concluded the eNOS is an AKT substrate linking signal transduction by AKT to the release of the gaseous second messenger nitric oxide. AKT mediates the activation of eNOS, leading to increased nitric oxide production. Inhibition of the PI3K AKT pathway or mutation of the AKT site on eNOS protein at serine-1177 attenuated the serine phosphorylation and prevented the activation of eNOS. RT-PCR analysis showed that expression ofNOS3in human umbilical vein endothelial cells (HUVECs) and human aortic vascular smooth muscle cells (HAOVSMCs) was inversely proportional to that of NOS3AS.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5 µg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 µg/ml, Human
Flow Cytometry, 1-3 µg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5 µg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of eNOS/NOS3 using anti-eNOS/NOS3 antibody (A01604-3).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human placenta tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-eNOS/NOS3 antigen affinity purified polyclonal antibody (Catalog # A01604-3) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for eNOS/NOS3 at approximately 140 kDa. The expected band size for eNOS/NOS3 is at 133 kDa.

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Figure 2. IF analysis of eNOS/NOS3 using anti-eNOS/NOS3 antibody (A01604-3).
eNOS/NOS3 was detected in an immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-eNOS/NOS3 Antibody (A01604-3) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Figure 3. Flow Cytometry analysis of U937 cells using anti-eNOS/NOS3 antibody (A01604-3).
Overlay histogram showing U937 cells stained with A01604-3 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-eNOS/NOS3 Antibody (A01604-3, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For NOS3 (Source: Uniprot.org, NCBI)

Alternative Names

Constitutive NOS; EC 1.14.13.39; ECNOS; Endothelial NOS; endothelial; eNOS; eNOScNOS; nitric oxide synthase 3 (endothelial cell); NOS type III; NOS3; NOSIII NOS3 ECNOS, eNOS nitric oxide synthase 3 nitric oxide synthase, endothelial|EC-NOS|NOS type III|NOSIII|cNOS|constitutive NOS|endothelial NOS|nitric oxide synthase 3 (endothelial cell)|nitric oxide synthase 3 transcript variant eNOS-delta20|nitric oxide synthase 3 transcript variant eNOS-delta20-21|nitric oxide synthase 3 transcript variant eNOS-delta21

For more info on NOS3, check out the NOS3 Infographic

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In this infographic, you will see the following information for NOS3: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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