Anti-Nitro tyrosine antibody [HM.11] (ab7048)

Reviews (13)Q&A (21)References (38)

Overview

  • Product name
    Anti-Nitro tyrosine antibody [HM.11]
    See all Nitro tyrosine primary antibodies
  • Description
    Mouse monoclonal [HM.11] to Nitro tyrosine
  • Host species
    Mouse
  • Specificity
    This antibody reacts with nitrotyrosine, both with the free amino acid as well as with proteins containing nitrotyrosine. The antibody does not recognize other antigens with a nitrotyrosine-like structure such as tyrosine, 3-chlorotyrosine, phosphotyrosine and phenylalanine.
  • Tested applications
    Suitable for: WB, ICC/IF, ELISA, IHC-P, IHC-Frmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic peptide corresponding to Nitro tyrosine (nitrated ) conjugated to Keyhole Limpet Haemocyanin (KLH).

  • Positive control
    • WB: mouse kidney lysate, mouse optic nerve, retina , spinal cord and brain lysates, rat aorta lysate(see Abreviews) IHC-P: human lung tissue (see Abreview)

Properties

Applications

Our Abpromise guarantee covers the use of ab7048 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration. PubMed: 22462810
ELISA Use at an assay dependent concentration.
IHC-P 1/200 - 1/400. 10% formalin fixation; 3% H2O2 to block endogenous peroxidases; Citrate buffer pH 6.0 for 1 min at 100°C as antigen retrieval treatment
IHC-Fr Use at an assay dependent concentration. Acetone fixation 10 min -20°C; block endogenous peroxidase by 0.3 % H2O2 in PBS (or methanol for intracellular staining); block with 10% NGS or 5 % BSA for 30 min.

Target

  • Relevance
    Nitric oxide (NO) is a product of the enzymatic conversion of arginine to citrulline by nitric oxide synthase. NO reacts rapidly with superoxide to form peroxynitrite. At physiological pH and in the presence of transition metals, peroxynitrite undergoes heterolytic cleavage to form hydroxyl anion and nitronium ion, the latter of which nitrates protein tyrosine residues. Thus, the presence of nitrotyrosine on proteins can be used as a marker for peroxynitrite formation in vivo. The presence of nitrotyrosine has been detected in various inflammatory processes including atherosclerotic placques.
  • Alternative names
    • NO tyrosine antibody
    • nTyr antibody

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-Nitro tyrosine antibody [HM.11] (ab7048)
    Immunocytochemistry/ Immunofluorescence - Anti-Nitro tyrosine antibody [HM.11] (ab7048)Image from Laschak M et al., BMC Cancer. 2012 Mar 30;12:130. Fig 1.; doi:10.1186/1471-2407-12-130; 30 March 2012, BMC Cancer 2012, 12:130
    Immunofluorescence analysis of 22Rv1 cells (Human prostate carcinoma epithelial cell line) treated with JS-K, staining Nitro tyrosine (green) with ab7048.

    Cells were fixed with paraformaldehyde, permeabilized in 0.2% Triton X-100 and blocked with 1% BSA. Cells were incubated with primary antibody (1/25) for 30 min at 37°C. An AlexaFluor488®-conjugated anti-mouse IgG (1/100) was used as the secondary antibody.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nitro tyrosine antibody [HM.11] (ab7048)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nitro tyrosine antibody [HM.11] (ab7048)
    ab7048 at 2 µg/ml staining Nitrotyrosine in Human liver from a severely obese patient. ab7048 was incubated for 1 hour at room temperature.

References

This product has been referenced in:
  • Auger C  et al. Metformin adapts its cellular effects to bioenergetic status in a model of metabolic dysfunction. Sci Rep 8:5646 (2018). Read more (PubMed: 29618839) »
  • Zhang J  et al. Repeated superovulation increases the risk of osteoporosis and cardiovascular diseases by accelerating ovarian aging in mice. Aging (Albany NY) 10:1089-1102 (2018). Read more (PubMed: 29787998) »
See all 38 Publications for this product

Publishing research using ab7048? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 34 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Nitro tyrosine antibody [HM.11]

 Inconclusive
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rhesus monkey Tissue sections (Colon)
Specification
Colon
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer
Permeabilization
Yes - 10 mM Tris-HCl pH-8.0, 150 mM NaCl containing 0.1% Triton-X-100
Blocking step
3%BSA and 10% normal goat serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
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Abcam user community

Verified customer

Submitted Apr 11 2013

Question

danke für Ihre schnelle Antwort. Angehängt schicke ich Ihnen den Fragebogen. Die Problematik die wir sehen steht auch drinnen beschrieben. Wir hoffen wir können zu einer Lösung unseres Problem kommen.

Mit freundlichen Grüßen,

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Answer

Vielen Dank für Ihre Anfrage und dafür, dass Sie sich die Zeit genommen haben, unseren Fragebogen auszufüllen.

Es tut mir leid, dass Sie Probleme mit unserem Antikörper hatten.

Um Ihr Problem zu lösen, möchte ich die folgenden Vorschläge zu Veränderung Ihres Protokolls machen:

1. Um die Qualität des Antikörpers zu überprüfen, wäre es hilfreich eine positiv Kontrolle durchzuführen. Als positiv Kontrollen für Mausproben empfehlen wir auf unserem Datenblatt beispielsweise mouse kidney lysate, mouse optic nerve, retina , spinal cord and brain lysates.

2. Um die Qualität der Proben zu testen würden wir empfehlen eine Ladekontrolle in das Experiment zu integrieren. Dieses würde zusaetzlich zeigen, dass die Proteinproben auf die Membran übertragen werden.

3. Da die detektierten Banden in den Proben von der Größe mit der schweren und leichten Kette von Antikörpern übereinstimmen, ist zu vermuten das endogene in der Probe enthaltene Antikörper von den verwendeten sekundären Antikörpern erkannt werden. Diese kann durch die Verwendung eines Antikörpers vermieden werden, der nur die native Form des IgG erkennt. Einen solchen können Sie in unserem Katalog finden (Anti-mouse IgG VeriBlot for IP secondary antibody (HRP) (ab131368) ).


Bitte lassen Sie mich wissen, ob ich Ihnen helfen konnte und zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

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Question

We currently don't have nitrated BSA (ab131379) from Abcam. We have a nitrated BSA from Sigma which could not be recognized by ab7048, and I was told the nitration percentage of the product is not very high. We are thinking about getting the nitrated BSA from Abcam. But before I place the order, my supervisor wants some guarantee that nitrated BSA (ab131379) will work with the ab7048 antibody, and if it doesn't, can we have a replacement or reimpurse?

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Answer



There is a western blot on our online datasheet for ab131379 https://www.abcam.com/bovine-serum-albumin-protein-nitrated-ab131379.html illustrating this protein to be detectable by ab110282, another Anti-Nitrotyrosine antibody. So we will guarantee that ab131379 will be a good positive control for western blotting, as long as you follow up with us within 6 months of purchasing the product.

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Question

herzlichen Dank für Ihre mail.

Nach Rücksprache mit Fr. Dr. Deindl würden wir gerne den ab7048 Anti-Nitro -tyrosineantibody ( 50µg) bestellen und testen für IP.

Zusätzlich möchten wir

Anti-SOD2 ab 13533 50µg ( 410,00€)

Anti-p53 antibody ab 90363 50µg (370,00€)

Anti-IKB alpha antibody ab 32518 100µg ( 410,00€) Rabmab ( Promotion-Code RabMab-XBSMF)

bestellen.

Könnten Sie uns bitte ein Angebot für die Antikörper machen einschließlich Rabmab ( + Versandkosten) und bitte Angebotsnumer?

Wir würden gerne auch noch Sepharose ( Protein A oder Protein G) für die Präzipitation bestellen. Könnten Sie uns auch hierfür ein Angebot machen?
Wir arbeiten mit Maus Gewebe ( Kollateralen und Adduktor).

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Answer



Es freut mich zu hören, dass Sie unser Angebot unseren Antikörper ab7048 in IP zu testen, annehmen möchten. Wie am Telefon besprochen, können Sie den Gutscheincode zum Kauf eines weiteren Antikörpers einsetzen, sobald Sie uns ein Abreview für diesen Antikörper eingereicht haben. Der Discountcode deckt den Wert von ab7048 ] ab und kann zum Kauf weiterer Produkte eingesetzt werden.

Der Code wird gültig, sobald Sie uns ein Abreview für diesen Antikörper über den Test in IP eingereicht haben. Bitte geben Sie diesen Code auch im Abschnitt “Additional Notes” des Abreviews mit an, so dass wir wissen, dass sich dieses Abreview auf die Gutscheinaktion bezieht. Der Code wird dadurch aktiviert. Bitte kontaktieren Sie bei der nächsten Bestellung unsere Kundendienst mit dem Code und der ursprünglichen Bestellnummer.

Bitte zögern Sie nicht, sich wieder an uns zu wenden, falls Sie weitere Fragen haben. Wir freuen uns auf Ihr Abreview, egal ob Ihre Ergebnisse positiv oder negativ sind und wünschen Ihnen viel Glück für Ihre Forschung.

Die Bedingungen dieses Angebotes befinden sich unter dem folgenden Link: www.abcam.com/collaborationdiscount


Leider haben wir keine Protein A/G Beads in unserem Katalog, so dass ich ihnen hierzu kein Angebot machen kann. Desweiteren habe ich eine Email an meine Kolleginnen vom Kundendienst weitergeleitet.

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Question

Will this Anti-Nitro tyrosine antibody [HM.11] (ab7048) work in IP?

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Answer

Vielen Dank für Ihren Anruf und für Ihr Interesse an unseren Produkten.

Unseres Wissens nach wurde Anti-Nitro tyrosine antibody [HM.11] (ab7048) bisher noch nicht in IP getestet. Falls Sie dies selbst testen möchten, kann ich Ihnen zur Zeit ein spezielles Angebot über einen 100%igen Abreview-Rabatt anbieten. Bei diesem Angebot bekommen Sie einen Rabatt für eine zukünftige Bestellung, wenn Sie uns ein Abreview mit dem Testresultat zusenden. Der Rabatt würde den ganzen Wert von ab7048abdecken, und gegen eine erneute Bestellung eines weiteren Produktes von uns verrechnet werden.

Um von diesem Angebot profitieren zu können, folgen Sie bitte diesen Schritten:

1.) Bestätigen Sie mir bitte, dass Sie ab7048 in IP testen möchten.

2.) Bitte bestellen Sie erst nach Erhalt des Rabattcodes.

3.) Bestellen Sie den Antikörper wie üblich per Telefon, Email oder Fax

4.) Testen Sie den Antikörper in IP.

5.) Teilen Sie uns das Ergebnis Ihres Tests durch ein Abreview mit und notieren Sie den Discount Code in dem Feld "additional notes"
Unter der folgenden URL können Sie mehr über unser Abreview System erfahren:
https://www.abcam.com/abreviews

6.) Der Rabattcode ist nach dem Abschicken des Abreviews aktiv. Bitte beachten Sie, dass der Rabattcode innerhalb von 4 Monaten nach Ausstellung eingelöst werden muss.

Der Rabattcode wird gültig unabhängig davon, ob Ihr Ergebnis positiv oder negativ ist.

Die Bedingungen zu unserem 100% Abreview Rabatt können Sie unter dem folgenden Link nachlesen:
https://www.abcam.com/collaborationdiscount

Ich hoffe, diese Informationen helfen Ihnen weiter. Falls Sie einen Rabattcode erhalten möchten oder weitere Fragen haben, zögern Sie bitte nicht, sich wieder an mich zu wenden.

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Question

Is nitrated BSA, such as ab131379, a good control for this antibody in western blotting?

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Answer

Our nitrated BSA (ab131379) should be a good control for use with ab7048.

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Question

Thank you for taking the time to help us out with this antibody. We contacted another lab who is successfully using another one of your antibodies to nitrotyrosine, specifically ab78163. We would be happy to get a refund and place an order for that antibody, or just get that antibody sent to us as a replacement. Whichever route is the easiest is fine with me. Would you like us to send back the remaining nitrotyrosine antibody that isn't working for us?
The original order number for the ab7048 antibody was **


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Answer

Thank you for calling Abcam earlier today.

I am sorry about the issues you have been having with ab7048. Asrequested I am sending you a new vial of the antibody, ab78163and you should receive that tomorrow morning. For your reference, your new order number is **.

If there is anything else I can help you with, please let me know.

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Question

I have attached our current protocol for staining HUVEC cells. The secondary antibody I have been using with ab7048 is a donkey anti-mouse Alexa 594. I have confirmed that the secondary antibody works using an independent antibody for actin.

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Answer

Thank you for providing that information.

Having looked over the protocol you provided, I do not think that there is any advice I could give that would help resolve the issues that you are having. Therefore I would be happy o either replace or refund the cost of the antibody for you.

Please let me know how you would you like to continue and if you could also provide either the Abcam order number or the PO# used to purchase the antibody, that would be very helpful.

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Question

Hello,
We recently purchased the Anti-Nitro tyrosine mouse monoclonal (HM.11) that was listed as ab7048. I can no longer find this catalog number in your system, and I am assuming that ab94883 is the same product. Our problem is that we detect absolutely no staining using this antibody. We are staining the HUVEC cell line, and human endothelial cells collections. Both of these have been fixed (0.75% Formaldehyde OR 4% Paraformaldehyde), frozen and affixed to slides. We purchased this antibody because my PI's previous lab had successful staining on HUVECs with it, and we are using their protocol. When we co-stain with ab8822 (Anti-Von Willebrand Factor (FITC)), we get a beautiful signal from ab8822 and nothing from ab7048. When we stain with ab7048 alone, we likewise get nothing. We have even treated the cells with ROS prior to fixing them to induce a response. We are using Donkey anti-mouse Alexafluor 596 as a secondary antibody. We have confirmed that this secondary antibody is working by using it with a separate primary antibody to actin. Do you have any suggestions to get this antibody to work? Alternatively, we would be happy to trade it in for a different nitro tyrosine antibody if this one is problematic.

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Answer

Thank you for contacting Abcam.

I am sorry about the problems you have been having with ab7048, which is still in the catalogue and can be found at the link below:

https://www.abcam.com/Nitro-tyrosine-antibody-HM-11-ab7048.html



The antibody is covered under our Abpromise for six months and is guaranteed to work inIHC-P& IHC-Fr. If we cannot resolve the issue you are having with the antibody then I would be happy to either send a replacement antibody or to process a refund.

To be able to help you further in this matter would you be able to send me the complete protocol that you are using. I know you mentioned that you were getting really good staining with the protocol previously and that it worked for ab8822, but I may be able to make some suggestions. If not, then I will be happy to replace the antibody for you.

I look forward to your reply and helping to resolve this issue.

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Question

Very high background (membrane is blotchy) and lots of non-specific bands.

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Answer

Thank you for calling Abcam earlier today.

As we discussed, I am sending you a new vial of antibody, ab42789 as a free of charge replacement. Your new order number is *********.

The antibody is covered under our Abpromise to work in western blot and the Abpromise is valid for 6 months after delivery. If you have any issues with this antibody, please contact us within that time frame to let us know.

I hope that your experiments go well.

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