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Product Info Summary
| SKU: | A01604-2 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-eNOS/NOS3 Antibody Picoband®
SKU/Catalog Number
A01604-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-eNOS/NOS3 Antibody Picoband® catalog # A01604-2. Tested in ELISA, Flow Cytometry, IHC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store 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 freeze-thaw cycles.
Cite This Product
Anti-eNOS/NOS3 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01604-2)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human eNOS/NOS3 recombinant protein (Position: Q1052-P1203).
*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
A01604-2 is reactive to NOS3 in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
133 kDa
Calculated molecular weight
149527 MW
Background of eNOS
NOS3 (Nitric Oxide Synthase 3), also called ENOS, a nitric oxide synthase that generates NO in blood vessels and is involved with regulating vascular tone by inhibiting smooth muscle contraction and platelet aggregation. The NOS3 gene is mapped on 7q36.1. Variations in this gene are associated with susceptibility tocoronary spasm. Fulton et al. 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 of NOS3 in 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.
Application & Images
Applications
A01604-2 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee
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, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat, By Heat
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, Huma
Positive Control
WB: human placenta tissue, human K562 whole cell, human HepG2 whole cell, human THP-1 whole cell, rat kidney tissue, mouse kidne tissue, mouse small intestine tissue,
IHC: human lung cancer tissue, rat spleen tissue, mouse spleen tissue
FCM: HepG2 cell
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of NOS3 using anti-NOS3 antibody (A01604-2).
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 50ug of sample under reducing conditions.
Lane 1: human placenta tissue lysates
Lane 2: human K562 whole cell lysates
Lane 3: human HepG2 whole cell lysates
Lane 4: human THP-1 whole cell lysates
Lane 5: rat kidney tissue lysates
Lane 6: mouse kidne tissue lysates
Lane 7: mouse small intestine tissue lysates
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-NOS3 antigen affinity purified polyclonal antibody (Catalog # A01604-2) 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 1:10000 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 NOS3 at approximately 133KD. The expected band size for NOS3 is at 133KD.
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Figure 2. IHC analysis of NOS3 using anti-NOS3 antibody (A01604-2).
NOS3 was detected in paraffin-embedded section of human lung cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-NOS3 Antibody (A01604-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 3. IHC analysis of NOS3 using anti-NOS3 antibody (A01604-2).
NOS3 was detected in paraffin-embedded section of rat spleen tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-NOS3 Antibody (A01604-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 4. IHC analysis of NOS3 using anti-NOS3 antibody (A01604-2).
NOS3 was detected in paraffin-embedded section of mouse spleen tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-NOS3 Antibody (A01604-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 5. Flow Cytometry analysis of HepG2 cells using anti-NOS3 antibody (A01604-2).
Overlay histogram showing HepG2 cells stained with A01604-2 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NOS3 Antibody (A01604-2,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Protein Target Info & Infographic
Gene/Protein Information For NOS3 (Source: Uniprot.org, NCBI)
Gene Name
NOS3
Full Name
Nitric oxide synthase, endothelial
Weight
149527 MW
Superfamily
NOS family
Alternative Names
Nitric oxide synthase, endothelial; Constitutive NOS; Cnos; EC-NOS; Endothelial NOS; eNOS; NOS type III; NOSIII; NOS3 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
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on NOS3, check out the NOS3 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
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].
Specific Publications For Anti-eNOS/NOS3 Antibody Picoband® (A01604-2)
Hello CJ!
A01604-2 has been cited in 23 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
Inhibition by brimonidine of forskolin-induced nitric oxide synthase expression in human ciliary bodies in vitro
Propofol improves cardiac functional recovery after ischemia-reperfusion by upregulating nitric oxide synthase activity in the isolated rat hearts
Research on the Age-Related Changes in the Nitric Oxide Pathway in the Arteries of Rats and the Intervention Effect of Dehydroepiandrosterone
Effects of Quercetin on Gene and Protein Expression of NOX and NOS after Myocardial Ischemia and Reperfusion in Rabbit
Fucoidan from Undaria pinnatifida prevents vascular dysfunction through PI3K/Akt/eNOS-dependent mechanisms in the L-NAME-induced hypertensive rat model
Effects of Calcitonin Gene-Related Peptide on the Expression and Activity of Nitric Oxide Synthase During Mandibular Bone Healing in Rabbits: An Experimental Study
Sesamin improves endothelial dysfunction in renovascular hypertensive rats fed with a high-fat, high-sucrose diet
Protective effects of 2,3,5,4′-tetrahydroxystilbene-2-O-beta-d-glucoside, an active component of Polygonum multiflorum Thunb, on experimental colitis in mice
Increased secretion of VEGF-C from SiO2-induced pulmonary macrophages promotes lymphangiogenesis through the Src/eNOS pathway in silicosis
Autologous Bone Marrow Mononuclear Cell Transplantation Delays Progression of Carotid Atherosclerosis in Rabbits
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Customer Q&As
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16 Customer Q&As for Anti-eNOS/NOS3 Antibody Picoband®
Question
Does anti-eNOS/NOS3 antibody A01604-2 work for ELISA with umbilical vein?
Verified Customer
Verified customer
Asked: 2020-03-17
Answer
According to the expression profile of umbilical vein, NOS3 is highly expressed in umbilical vein. So, it is likely that anti-eNOS/NOS3 antibody A01604-2 will work for ELISA with umbilical vein.
Boster Scientific Support
Answered: 2020-03-17
Question
Do you have a BSA free version of anti-eNOS/NOS3 antibody A01604-2 available?
Verified Customer
Verified customer
Asked: 2020-02-19
Answer
Thanks for your recent telephone inquiry. I can confirm that some lots of this anti-eNOS/NOS3 antibody A01604-2 are BSA free. For now, these lots are available and we can make a BSA free formula for you free of charge. It will take 3 extra days to prepare. If you require this antibody BSA free again in future, please do not hesitate to contact me and I will be pleased to check which lots we have in stock that are BSA free.
Boster Scientific Support
Answered: 2020-02-19
Question
Is a blocking peptide available for product anti-eNOS/NOS3 antibody (A01604-2)?
Verified Customer
Verified customer
Asked: 2019-12-23
Answer
We do provide the blocking peptide for product anti-eNOS/NOS3 antibody (A01604-2). If you would like to place an order for it please contact [email protected] and make a special request.
Boster Scientific Support
Answered: 2019-12-23
Question
We are currently using anti-eNOS/NOS3 antibody A01604-2 for mouse tissue, and we are happy with the IHC-P results. The species of reactivity given in the datasheet says human, mouse, rat. Is it likely that the antibody can work on canine tissues as well?
Verified Customer
Verified customer
Asked: 2019-10-11
Answer
The anti-eNOS/NOS3 antibody (A01604-2) has not been tested for cross reactivity specifically with canine tissues, though there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in canine you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-10-11
Question
I appreciate helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for umbilical vein using anti-eNOS/NOS3 antibody A01604-2. Let me know if you need anything else.
Verified Customer
Verified customer
Asked: 2019-04-16
Answer
We appreciate the data. You have provided everything we needed. Our lab team are working to resolve your inquiry as quickly as possible, and we appreciate your patience and understanding! Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2019-04-16
Question
I am interested in using your anti-eNOS/NOS3 antibody for positive regulation of blood vessel endothelial cell migration studies. Has this antibody been tested with western blotting on small intestine tissue? We would like to see some validation images before ordering.
Verified Customer
Verified customer
Asked: 2019-04-03
Answer
We appreciate your inquiry. This A01604-2 anti-eNOS/NOS3 antibody is tested on human placenta tissue, k562 whole cell lysates, hepg2 whole cell lysates, rat spleen tissue, kidney tissue, mouse kidne tissue lysates, small intestine tissue, u87 cells. It is guaranteed to work for ELISA, Flow Cytometry, IHC-P, WB in human, mouse, rat. Our Boster guarantee will cover your intended experiment even if the sample type has not been be directly tested.
Boster Scientific Support
Answered: 2019-04-03
Question
Our team were happy with the WB result of your anti-eNOS/NOS3 antibody. However we have observed positive staining in spleen caveola. cytoplasm, using this antibody. Is that expected? Could you tell me where is NOS3 supposed to be expressed?
Verified Customer
Verified customer
Asked: 2018-10-30
Answer
From literature, spleen does express NOS3. Generally NOS3 expresses in cell membrane. membrane, caveola. cytoplasm,. Regarding which tissues have NOS3 expression, here are a few articles citing expression in various tissues:
Erythroleukemia, Pubmed ID: 23186163
Hippocampus, and Trachea, Pubmed ID: 14702039
Liver, Pubmed ID: 24275569
Placenta, Pubmed ID: 7509596, 7514568, 7515611, 15489334
Platelet, Pubmed ID: 7475956
Spleen, Pubmed ID: 12853948
Boster Scientific Support
Answered: 2018-10-30
Question
I have attached the WB image, lot number and protocol we used for umbilical vein using anti-eNOS/NOS3 antibody A01604-2. Please let me know if you require anything else.
Verified Customer
Verified customer
Asked: 2018-10-03
Answer
Thank you very much for the data. Our lab team are working to resolve this as quickly as possible, and we appreciate your patience and understanding! You have provided everything we needed. Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2018-10-03
Question
Does A01604-2 anti-eNOS/NOS3 antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
Verified Customer
Verified customer
Asked: 2018-05-15
Answer
It shows on the product datasheet, A01604-2 anti-eNOS/NOS3 antibody as been tested on ELISA. It is best to use PFA for fixation because it has better tissue penetration ability. PFA needs to be prepared fresh before use. Long term stored PFA turns into formalin, as the PFA molecules congregate and become formalin.
Boster Scientific Support
Answered: 2018-05-15
Question
We have observed staining in rat umbilical vein. What should we do? Is anti-eNOS/NOS3 antibody supposed to stain umbilical vein positively?
T. Miller
Verified customer
Asked: 2018-01-04
Answer
According to literature umbilical vein does express NOS3. According to Uniprot.org, NOS3 is expressed in spleen, placenta, umbilical vein, platelet, hippocampus trachea, erythroleukemia, liver, among other tissues. Regarding which tissues have NOS3 expression, here are a few articles citing expression in various tissues:
Erythroleukemia, Pubmed ID: 23186163
Hippocampus, and Trachea, Pubmed ID: 14702039
Liver, Pubmed ID: 24275569
Placenta, Pubmed ID: 7509596, 7514568, 7515611, 15489334
Platelet, Pubmed ID: 7475956
Spleen, Pubmed ID: 12853948
Boster Scientific Support
Answered: 2018-01-04
Question
I see that the anti-eNOS/NOS3 antibody A01604-2 works with ELISA, what is the protocol used to produce the result images on the product page?
Verified Customer
Verified customer
Asked: 2017-06-14
Answer
You can find protocols for ELISA on the "support/technical resources" section of our navigation menu. If you have any further questions, please send an email to [email protected]
Boster Scientific Support
Answered: 2017-06-14
Question
Is this A01604-2 anti-eNOS/NOS3 antibody reactive to the isotypes of NOS3?
S. Jha
Verified customer
Asked: 2016-03-11
Answer
The immunogen of A01604-2 anti-eNOS/NOS3 antibody is E.coli-derived human eNOS/NOS3 recombinant protein (Position: Q1052-P1203). Could you tell me which isotype you are interested in so I can help see if the immunogen is part of this isotype?
Boster Scientific Support
Answered: 2016-03-11
Question
I was wanting to use your anti-eNOS/NOS3 antibody for ELISA for rat umbilical vein on frozen tissues, but I want to know if it has been validated for this particular application. Has this antibody been validated and is this antibody a good choice for rat umbilical vein identification?
S. Parker
Verified customer
Asked: 2015-04-29
Answer
It shows on the product datasheet, A01604-2 anti-eNOS/NOS3 antibody has been validated for ELISA, Flow Cytometry, IHC-P, WB on human, mouse, rat tissues. We have an innovator award program that if you test this antibody and show it works in rat umbilical vein in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2015-04-29
Question
We want to test anti-eNOS/NOS3 antibody A01604-2 on rat umbilical vein for research purposes, then I may be interested in using anti-eNOS/NOS3 antibody A01604-2 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
A. Walker
Verified customer
Asked: 2014-12-15
Answer
The products we sell, including anti-eNOS/NOS3 antibody A01604-2, are only intended for research use. They would not be suitable for use in diagnostic work. If you have the means to develop a product into diagnostic use, and are interested in collaborating with us and develop our product into an IVD product, please contact us for more discussions.
Boster Scientific Support
Answered: 2014-12-15
Question
We bought anti-eNOS/NOS3 antibody for WB on hippocampus trachea in a previous experiment. I am using mouse, and I plan to use the antibody for IHC-P next. I am interested in examining hippocampus trachea as well as erythroleukemia in our next experiment. Could give a recommendation on which antibody would work the best for IHC-P?
S. Evans
Verified customer
Asked: 2013-05-15
Answer
I have checked the website and datasheets of our anti-eNOS/NOS3 antibody and I see that A01604-2 has been validated on mouse in both WB and IHC-P. Thus A01604-2 should work for your application. Our Boster satisfaction guarantee will cover this product for IHC-P in mouse even if the specific tissue type has not been validated. We do have a comprehensive range of products for IHC-P detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2013-05-15
Question
I have a question about product A01604-2, anti-eNOS/NOS3 antibody. I was wondering if it would be possible to conjugate this antibody with biotin. I would need it to be without BSA or sodium azide. I am planning on using a buffer exchange of sodium azide with PBS only. Would there be problems for me to conjugate the antibody and store it in -20 degrees in small aliquots?
R. Banerjee
Verified customer
Asked: 2013-04-29
Answer
It is not recommended storing this antibody with PBS buffer only in -20 degrees. If you want to store it in -20 degrees it is best to add some cryoprotectant like glycerol. If you want carrier free A01604-2 anti-eNOS/NOS3 antibody, we can provide it to you in a special formula with trehalose and/or glycerol. These molecules will not interfere with conjugation chemistry and provide a good level of protection for the antibody from degradation. Please be sure to specify this in your purchase order.
Boster Scientific Support
Answered: 2013-04-29
