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Product Info Summary
| SKU: | A00368 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | IHC, WB |
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Product info
Product Name
Anti-iNOS/NOS2 Antibody Picoband®
SKU/Catalog Number
A00368
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-iNOS/NOS2 Antibody Picoband® catalog # A00368. Tested in IHC, WB applications. This antibody reacts with Human. 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-iNOS/NOS2 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00368)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence at the C-terminus of human iNOS, different from the related mouse sequence by five amino acids, and from the related rat sequence by four amino acids.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00368 is reactive to NOS2 in Human
Observed Molecular Weight
131 kDa
Calculated molecular weight
131.1 kDa
Background of NOS2
Nitric oxide synthase, inducible is an enzyme that in humans is encoded by the NOS2 gene. Nitric oxide (NO) is a messenger molecule with diverse functions throughout the body. In the brain and peripheral nervous system, NO displays many properties of a neurotransmitter; it is implicated in neurotoxicity associated with stroke and neurodegenerative diseases, neural regulation of smooth muscle, including peristalsis, and penile erection. Three different NOS isoforms have been identified which fall into two distinct types, constitutive and inducible. The inducible NOS (iNOS) isoform is expressed in a variety of cell types and tissues in response to inflammatory agents and cytokines. The human iNOS (NOS2) gene is approximately 37 kb in length and consists of 26 exons and 25 introns. NOS2-derived NO is a prerequisite for cytokine signaling and function in innate immunity.
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
A00368 is guaranteed for 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
Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human
Positive Control
WB: human Hela whole cell, human Caco-2 whole cell, human SH-SY5Y whole cell
IHC: human brain tissue
Validation Images & Assay Conditions
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Western blot analysis of NOS2 using anti-NOS2 antibody (A00368).
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 Hela whole cell lysates,
Lane 2: human Caco-2 whole cell lysates,
Lane 3: human SH-SY5Y whole cell 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-NOS2 antigen affinity purified polyclonal antibody (Catalog # A00368) 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 NOS2 at approximately 131 kDa. The expected band size for NOS2 is at 131 kDa.
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IHC analysis of iNOS/NOS2 using anti-iNOS/NOS2 antibody (A00368).
iNOS/NOS2 was detected in a paraffin-embedded section of human brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-iNOS/NOS2 Antibody (A00368) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
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Celastrol inhibited inducible nitric oxide synthase (iNOS) expression and activation of NF-κB in optic nerve in EAE rats. (A) IHC staining of iNOS in optic nerve. (B) Quantification of iNOS-positive areas. (C) Quantitative real-time PCR analysis of iNOS expression. (D) Western blot analysis of iNOS expression. (E–G) Western blot analysis of IκBα, p65 and p-p65 expression, respectively. Treatment of celastrol reduced expression of iNOS and inhibited the activation of NF-κB in optic nerve in EAE rats. Scale bar: 100 μm. Data were shown as mean ± SD, n = 5. ∗∗ P < 0.01 versus control group, ## P < 0.01 versus EAE group, †† P < 0.01 versus low dosage of celastrol group.
Index in PubMed under a CC BY license. PMID: 28239352
Specific Publications For Anti-iNOS/NOS2 Antibody Picoband® (A00368)
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Customer Q&As
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15 Customer Q&As for Anti-iNOS/NOS2 Antibody Picoband®
Question
We want to test anti-iNOS/NOS2 antibody A00368 on human glioblastoma for research purposes, then I may be interested in using anti-iNOS/NOS2 antibody A00368 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
Verified Customer
Verified customer
Asked: 2020-05-01
Answer
The products we sell, including anti-iNOS/NOS2 antibody A00368, 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: 2020-05-01
Question
We have been able to see staining in human colon adenocarcinoma. Any tips? Is anti-iNOS/NOS2 antibody supposed to stain colon adenocarcinoma positively?
M. Anderson
Verified customer
Asked: 2020-04-27
Answer
Based on literature colon adenocarcinoma does express NOS2. Based on Uniprot.org, NOS2 is expressed in cartilage tissue, colon adenocarcinoma, liver, chondrocyte, articular chondrocyte, retina, glioblastoma, airway epithelium, cardiac myocyte, kidney, among other tissues. Regarding which tissues have NOS2 expression, here are a few articles citing expression in various tissues:
Airway epithelium, Pubmed ID: 7544004
Articular chondrocyte, Pubmed ID: 7522054
Cardiac myocyte, Pubmed ID: 9160867
Chondrocyte, Pubmed ID: 7504305
Colon adenocarcinoma, Pubmed ID: 7692964
Glioblastoma, Pubmed ID: 7528267, 7531687
Kidney, Pubmed ID: 7532248
Liver, Pubmed ID: 7682706
Retina, Pubmed ID: 7528017
Boster Scientific Support
Answered: 2020-04-27
Question
I have attached the WB image, lot number and protocol we used for glioblastoma using anti-iNOS/NOS2 antibody A00368. Please let me know if you require anything else.
Verified Customer
Verified customer
Asked: 2020-04-06
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: 2020-04-06
Question
Will anti-iNOS/NOS2 antibody A00368 work for WB with glioblastoma?
Verified Customer
Verified customer
Asked: 2020-01-06
Answer
According to the expression profile of glioblastoma, NOS2 is highly expressed in glioblastoma. So, it is likely that anti-iNOS/NOS2 antibody A00368 will work for WB with glioblastoma.
Boster Scientific Support
Answered: 2020-01-06
Question
Can you help my question with product A00368, anti-iNOS/NOS2 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?
Verified Customer
Verified customer
Asked: 2019-12-16
Answer
We do not advise 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 A00368 anti-iNOS/NOS2 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: 2019-12-16
Question
Would A00368 anti-iNOS/NOS2 antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
Verified Customer
Verified customer
Asked: 2019-10-14
Answer
You can see on the product datasheet, A00368 anti-iNOS/NOS2 antibody as been tested on WB. 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: 2019-10-14
Question
Is a blocking peptide available for product anti-iNOS/NOS2 antibody (A00368)?
Verified Customer
Verified customer
Asked: 2019-10-03
Answer
We do provide the blocking peptide for product anti-iNOS/NOS2 antibody (A00368). If you would like to place an order for it please contact support@bosterbio.com and make a special request.
Boster Scientific Support
Answered: 2019-10-03
Question
Is this A00368 anti-iNOS/NOS2 antibody reactive to the isotypes of NOS2?
Verified Customer
Verified customer
Asked: 2019-08-30
Answer
The immunogen of A00368 anti-iNOS/NOS2 antibody is A synthetic peptide corresponding to a sequence at the C-terminus of human iNOS (1088-1126aa ARDVAHTLKQLVAAKLKLNEEQVEDYFFQLKSQKRYHED), different from the related mouse sequence by five amino acids, and from the related rat sequence by four amino acids. 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: 2019-08-30
Question
I see that the anti-iNOS/NOS2 antibody A00368 works with WB, what is the protocol used to produce the result images on the product page?
Verified Customer
Verified customer
Asked: 2019-06-03
Answer
You can find protocols for WB on the "support/technical resources" section of our navigation menu. If you have any further questions, please send an email to support@bosterbio.com
Boster Scientific Support
Answered: 2019-06-03
Question
Thank you for helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for glioblastoma using anti-iNOS/NOS2 antibody A00368. Let me know if you need anything else.
Verified Customer
Verified customer
Asked: 2019-01-17
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: 2019-01-17
Question
My boss were content with the WB result of your anti-iNOS/NOS2 antibody. However we have observed positive staining in cartilage tissue cytoplasm using this antibody. Is that expected? Could you tell me where is NOS2 supposed to be expressed?
Verified Customer
Verified customer
Asked: 2018-01-31
Answer
From literature, cartilage tissue does express NOS2. Generally NOS2 expresses in cytoplasm, cytosol. Regarding which tissues have NOS2 expression, here are a few articles citing expression in various tissues:
Airway epithelium, Pubmed ID: 7544004
Articular chondrocyte, Pubmed ID: 7522054
Cardiac myocyte, Pubmed ID: 9160867
Chondrocyte, Pubmed ID: 7504305
Colon adenocarcinoma, Pubmed ID: 7692964
Glioblastoma, Pubmed ID: 7528267, 7531687
Kidney, Pubmed ID: 7532248
Liver, Pubmed ID: 7682706
Retina, Pubmed ID: 7528017
Boster Scientific Support
Answered: 2018-01-31
Question
I was wanting to use using your anti-iNOS/NOS2 antibody for nitric oxide mediated signal transduction studies. Has this antibody been tested with western blotting on hela whole cell lysates? We would like to see some validation images before ordering.
Verified Customer
Verified customer
Asked: 2017-09-25
Answer
Thanks for your inquiry. This A00368 anti-iNOS/NOS2 antibody is validated on hela whole cell lysates. It is guaranteed to work for WB in human. Our Boster guarantee will cover your intended experiment even if the sample type has not been be directly tested.
Boster Scientific Support
Answered: 2017-09-25
Question
We are currently using anti-iNOS/NOS2 antibody A00368 for human tissue, and we are satisfied with the WB results. The species of reactivity given in the datasheet says human. Is it likely that the antibody can work on horse tissues as well?
N. Dhar
Verified customer
Asked: 2016-12-30
Answer
The anti-iNOS/NOS2 antibody (A00368) has not been validated for cross reactivity specifically with horse 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 horse you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2016-12-30
Question
Is there a BSA free version of anti-iNOS/NOS2 antibody A00368 available?
J. Zhang
Verified customer
Asked: 2014-05-02
Answer
We appreciate your recent telephone inquiry. I can confirm that some lots of this anti-iNOS/NOS2 antibody A00368 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: 2014-05-02
Question
I was wanting to use your anti-iNOS/NOS2 antibody for WB for human glioblastoma 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 human glioblastoma identification?
T. Kulkarni
Verified customer
Asked: 2013-10-25
Answer
You can see on the product datasheet, A00368 anti-iNOS/NOS2 antibody has been validated for WB on human tissues. We have an innovator award program that if you test this antibody and show it works in human glioblastoma in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2013-10-25
