Name: Anti-3-Nitrotyrosine antibody [39B6]
SKU: ab61392
Rating: 4 (9 reviews)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-3-Nitrotyrosine antibody [39B6] (ab61392)

Key features and details

Mouse monoclonal [39B6] to 3-Nitrotyrosine
Suitable for: IHC-P, WB
Reacts with: Species independent
Isotype: IgG2a

Biotin FITC HRP

製品名

Anti-3-Nitrotyrosine antibody [39B6]
3-Nitrotyrosine 一次抗体製品一覧
製品の詳細

Mouse monoclonal [39B6] to 3-Nitrotyrosine
由来種

Mouse
特異性

No detectable crossreactivity with non-nitrated tyrosine. Not species specific.
アプリケーション

適用あり: IHC-P, WBmore details
種交差性

交差種: Species independent
免疫原

Full length protein corresponding to 3-Nitrotyrosine. 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin
ポジティブ･コントロール
- SIN-1 (3-morpholinosydnonimine) treated BSA. Use between 1-2.5mM SIN-1 for 1-24 hours, grow cells under normal growth conditions. Please see Yasuda S, Idell S, Liu MC. Biochem J. 2007 Jan 15;401(2):497-503. PMID: 17002600. IHC-P: Mouse back skin and inflammed colon tissue; Human colon carcinoma tissue. Rat liver tissue. WB: A549 cell lysate.
特記事項

The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

製品の状態

Liquid
保存方法

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
バッファー

Preservative: 0.09% Sodium azide
Constituents: PBS, 50% Glycerol
Concentration information loading...
精製度

Protein G purified
ポリ/モノ

モノクローナル
クローン名

39B6
アイソタイプ

IgG2a
軽鎖の種類

kappa
研究分野

Compatible Secondaries
- Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
Corresponding Antibody
Isotype control
- Mouse IgG2a, kappa monoclonal [MG2a-53] - Isotype control (ab18415)
Related Products
- Tyrosine Assay Kit (Colorimetric) (ab185435)

The Abpromise guarantee

Abpromise保証は、次のテスト済みアプリケーションにおけるab61392の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション	Abreviews	特記事項
IHC-P	(3)	1/100.
WB	(6)	1/1000. We would recommend a 1:1000 dilution for the antibody, for 2 hours at RT. For blocking, use milk.

特記事項
IHC-P 1/100.
WB 1/1000. We would recommend a 1:1000 dilution for the antibody, for 2 hours at RT. For blocking, use milk.

関連性

Protein tyrosine nitration results in a post-translational modification that is increasingly receiving attention as an important component of nitric oxide signaling. While multiple nonenzymatic mechanisms are known to be capable of producing nitrated tyrosine residues, most tyrosine nitration events involve catalysis by metalloproteins such as myeloperoxidase, eosinophilperoxidase, myoglobin, the cytochrome P-450s, superoxide dismutase and prostacyclin synthase. Various studies have shown that protein tyrosinenitration is limited to specific proteins and that the process is selective. For example, exposure of human surfactant protein A, SP-A, to oxygen-nitrogen intermediates generated by activated alveolar macrophages resulted in specific nitration of SP-A at tyrosines 164 and 166, while addition of 1.2 mMCO 2 resulted in additional nitration at tyrosine 161. The presence of nitrotyrosine-containing proteins has shown high correlation to disease states such as atherosclerosis, Alzheimer’s disease, Parkinson’s disease and amyotrophic lateral sclerosis.
別名
- 3-Nitrotyrosine antibody
- 3-NT antibody
- 3NT antibody
- Nitrotyrosine antibody

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-3-Nitrotyrosine antibody [39B6] (ab61392)

ab61392 staining 3-nitrotyrosine in Bouin's fixed paraffin-embedded Rat liver tissue at 1/1000 dilution. FITC-conjugated goat anti-mouse IgG(green) was used as a secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-3-Nitrotyrosine antibody [39B6] (ab61392)

ab61392 staining 3-nitrotyrosine in Bouin's fixed paraffin-embedded backskin sections of transgenic mice.
Blocker: 5% goat serum in buffer. Buffer: 99% washing buffer (7g BSA and 222ul gelatin topped up to 1L dH2O), 1% heat inactivated goat serum. Primary Dilution: 1/100 for 1 hour at room temperature. Secondary: FITC-conjugated goat anti-mouse IgG, 1/50 for 1 hour at room temperature.
Western blot - Anti-3-Nitrotyrosine antibody [39B6] (ab61392)This image is courtesy of an anonymous Abreview

All lanes : Anti-3-Nitrotyrosine antibody [39B6] (ab61392) at 1/3000 dilution

All lanes : Mouse skeletal muscle

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Mouse HRP conjugated at 1/6000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Exposure time: 4 seconds

Blocked with 5% milk for 2 hours.
See Abreview
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-3-Nitrotyrosine antibody [39B6] (ab61392)

Formalin-fixed, paraffin-embedded mouse inflammed colon tissue stained for Nitrotyrosine using ab61392 at 1/1000000 dilution in immunohistochemical analysis.

Secondary Antibody: Biotin Goat Anti-Mouse at 1/2000 for 1 hour at RT. Counterstain: Mayer Hematoxylin (purple/blue) nuclear stain at 200 µl for 2 minutes at RT.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-3-Nitrotyrosine antibody [39B6] (ab61392)

Formalin-fixed, paraffin-embedded human colon carcinoma tissue stained for Nitrotyrosine using ab61392 at 1/25000 dilution in immunohistochemical analysis.

Secondary Antibody: Biotin Goat Anti-Mouse at 1/2000 for 1 hour at RT. Counterstain: Mayer Hematoxylin (purple/blue) nuclear stain at 200 µl for 2 minutes at RT.
Western blot - Anti-3-Nitrotyrosine antibody [39B6] (ab61392)

Anti-3-Nitrotyrosine antibody [39B6] (ab61392) at 1/1000 dilution (2.5 hours at RT with shaking) + A549 (Human lung carcinoma cell line) cell lysate at 15 µg

Secondary
Goat anti-mouse IgG HRP, 1 hour at RT with shaking at 1/1000 dilution

Block: 5% Skim milk powder in TBST.

Color Development: Chemiluminescent for HRP for 5 mins at RT.

Immunoprecipitation protocols
Immunohistochemistry protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

ab61392 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab61392 は 153 報の論文で使用されています。

Liu J et al. Oxidative-antioxidant imbalance in chronic sialadenitis of submandibular gland in human and rat. Oral Dis 29:1005-1016 (2023). PubMed: 34826194
Yu L et al. Fibroblast growth factor 10 ameliorates renal ischaemia-reperfusion injury by attenuating mitochondrial damage. Clin Exp Pharmacol Physiol 50:59-67 (2023). PubMed: 36111374
Lackovic J et al. Peroxynitrite Contributes to Behavioral Responses, Increased Trigeminal Excitability, and Changes in Mitochondrial Function in a Preclinical Model of Migraine. J Neurosci 43:1627-1642 (2023). PubMed: 36697259
Black AL et al. Sexual dimorphism in reactive oxygen species production and a role for integrin α1β1 in estrogen receptor α and β expression in articular cartilage. J Orthop Surg Res 18:170 (2023). PubMed: 36879303
Khayrullina G et al. Differential effects of NOX2 and NOX4 inhibition after rodent spinal cord injury. PLoS One 18:e0281045 (2023). PubMed: 36897852

View all Publications for this product

レビューと Q&A

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レビューを送る質問を送る

1-10 of 23 Abreviews or Q&A

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Mouse Tissue sections (Lung)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citrate Buffer

Permeabilization

Specification

Lung

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Yaser Salaymeh

Verified customer

投稿 Apr 20 2022

Application

Western blot

Sample

Human Cell lysate - whole cell (SHSY5Y)

Gel Running Conditions

Reduced Denaturing

Loading amount

30 µg

Specification

SHSY5Y

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Mar 10 2021

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Mouse Tissue sections (liver)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: 10 mM sodium citrate buffer (pH 6.0)

Permeabilization

Yes - 0.4% Triton X-100

Specification

liver

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Apr 18 2018

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Rabbit Tissue sections (Iliac vessel)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: citrate buffer

Permeabilization

Specification

Iliac vessel

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Feb 27 2017

Application

Western blot

Sample

Mouse Tissue lysate - whole (Skin)

Gel Running Conditions

Reduced Denaturing (10% Gel)

Loading amount

50 µg

Treatment

Cinnamaldehyde

Specification

Skin

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 18°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Aug 19 2016

Application

Western blot

Sample

Mouse Tissue lysate - whole (Skeletal muscle)

Gel Running Conditions

Reduced Denaturing (12%)

Loading amount

20 µg

Specification

Skeletal muscle

Blocking step

Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Feb 10 2016

Application

Western blot

Sample

Rat Tissue lysate - whole (Skeletal muscle)

Gel Running Conditions

Reduced Denaturing (12%)

Loading amount

20 µg

Specification

Skeletal muscle

Blocking step

Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Feb 09 2016

Application

Western blot

Loading amount

20 µg

Gel Running Conditions

Reduced Denaturing (12% gel)

Sample

Mouse Tissue lysate - whole (Skeletal muscle (GTN))

Specification

Skeletal muscle (GTN)

Blocking step

Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Jun 09 2014

Question

To whom it may concern, I had some problems with the ab61392 product in establishing, I get staining also in negative healthy controls, which I believe is background. On the other hand, my positive control (3-Morpholinosydnonimine (SIN-1) treated NT2 cells 1h 100µM or 1h 200µM) mostly seem to be similar to untreated NT2 cells, no good signal in both. I tried to troubleshoot a lot of steps incl. antigen retrieval, blocking, Ab dilutions, but I could not figure out why it is not working. I have almost run out of Ab. I now rather tend to think that it is the problem with the product. Another reason for this opinion, is the fact that I cannot be sure whether it is working or not. Sometimes I do see difference btw treated vs untreated cells, sometimes not. Even when I use 1mM SIN-1 for 4h, although most of the cells I see are shrunken and apoptotic, I do not see a better signal than it is in untreated control. and when I see signal I mostly see it from microtubules. I am totally exhausted and I would really appreciate any information regarding this issue. Additionally, I am afraid I should say none of the publications which used ab61392 -given in product page- was actually helpful nor convincing. Below are the latest conditions I use: Fixation: 4% Formalin Fixed and Paraffin Embedded Rat Brain Tissue Antigen Retrieval: 10mM Na-Citrate, pH6 20 min at 98°C Blocking: 0.5% BSA + 7% FBS in TBS 2h at room temperature. 1° Ab: 1,5µg/ml in blocking solution overnight incubation at 4°C 2° Ab Dilution: 2µg/ml in blocking solution Alexa 568TM conjugated 1h at room temperature. Finally, I would like to ask whether it is possible for you to send me a small aliquote from ab110282 for free. I would appreciate if that is possible and I would be happy to give a detailed feedback in the product page, and make an order from ab110282 if it works.

Abcam community

Verified customer

Asked on Apr 26 2013

Answer

The protocol you provided is for the IHC on rat brain, which you say showed some background staining but nothing that appeared specific. Did you have a treated/diseased rat brain for comparison? Did you include a negative control, using no primary antibody, to assess whether the background staining was contributed by the secondary antibody? Are your washes adequate for the thickness of the sections? I am assuming they are not thicker than 10um, but if they are much thicker, more washes may be required.

The ICC results that you describe are inconclusive: sometimes the antibody appeared to stain treated cells compared to untreated, and sometimes it did not, is that correct? Was there any correlation with length of SIN-1 treatment? Do you have any evidence, from the literature, when nitration in response to SIN-1 treatment is expected to be maximal? I am wondering if the 4h timepoint, when cells were apoptotic, was too long.

The positive control that is listed on the datasheet was detected with TLC, according to the Yasuda reference. We have no protocol for ICC/IF for this antibody. Can you please tell me how the cells were prepared for staining after treatment?

Abcam Scientific Support

Answered on Apr 26 2013

Question

Inquiry: We would like to test Anti-3-Nitrotyrosine Antibody in IHC-Fr of human skin.Do you have any references with this antibody on human skin? Thank you und kind regards

Abcam community

Verified customer

Asked on Aug 21 2012

Answer

Thank you for contacting us.

I am not aware of a reference using this antibody on human skin. On the datasheet you have an image of frozen section mouse skin. I am sorry that I can not help you more on this occasion.

Please be also reassured that we do guarantee all our products to work as stated on the datasheet. We do therefore also guarantee the ab61392 to work on human skin frozen sections.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Abcam Scientific Support

Answered on Aug 21 2012

1-10 of 23 Abreviews or Q&A

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Key features and details

Related conjugates and formulations

製品の概要

製品名

製品の詳細

由来種

特異性

アプリケーション

種交差性

免疫原

ポジティブ･コントロール

特記事項

製品の特性

製品の状態

保存方法

バッファー

精製度

ポリ/モノ

クローン名

アイソタイプ

軽鎖の種類

研究分野

関連製品

Compatible Secondaries

Corresponding Antibody

Isotype control

Related Products

アプリケーション

The Abpromise guarantee

ターゲット情報

関連性

別名

画像

プロトコール

データシートおよび資料

SDS download

Datasheet download

参考文献 (153)

レビューと Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-3-Nitrotyrosine antibody [39B6]

Western blot abreview for Anti-3-Nitrotyrosine antibody [39B6]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-3-Nitrotyrosine antibody [39B6]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-3-Nitrotyrosine antibody [39B6]

Western blot abreview for Anti-3-Nitrotyrosine antibody [39B6]

Western blot abreview for Anti-3-Nitrotyrosine antibody [39B6]

Western blot abreview for Anti-3-Nitrotyrosine antibody [39B6]

Western blot abreview for Anti-3-Nitrotyrosine antibody [39B6]