Overview

  • Product name
    Anti-Nitro tyrosine antibody [HM.11] (Biotin)
    See all Nitro tyrosine primary antibodies
  • Description
    Mouse monoclonal [HM.11] to Nitro tyrosine (Biotin)
  • Host species
    Mouse
  • Conjugation
    Biotin
  • Specificity
    This highly specific monoclonal antibody reacts with nitrotyrosine, both with the free amino acid as well as with proteins containing nitrotyrosine.
  • Tested applications
    Suitable for: WB, IHC-P, IHC-Frmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat

Properties

Applications

Our Abpromise guarantee covers the use of ab7225 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
AP Use at an assay dependent concentration.
WB Use at an assay dependent concentration.
IHC-P 1/200 - 1/400.
IHC-Fr Use at an assay dependent dilution.

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

  • Nitrotyrosine in human liver of severely obese patients. Staining of paraffin tissue section with ab7225 at 2 μg/ml.

References

This product has been referenced in:
  • Hathuc C  et al. Nitration of human plasminogen by RAW 264.7 macrophages reduces streptokinase-induced plasmin activity. Clin Chem Lab Med 44:213-9 (2006). WB . Read more (PubMed: 16475910) »
  • Bixby M  et al. Ilex paraguariensis extracts are potent inhibitors of nitrosative stress: a comparative study with green tea and wines using a protein nitration model and mammalian cell cytotoxicity. Life Sci 77:345-58 (2005). WB . Read more (PubMed: 15878361) »
See all 2 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Answer

Thank you for your inquiry. I would recommend to try a range of dilutions (1:1000- 1:20,000) for this antibody in Western blot. The optimal dilution needs to optimized according to the results.

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Question
Answer

These two references from the same lab suggest kidney carcinoma as a potential positive control for nitrotyrosine: Oxidative/nitrosative stress and peroxiredoxin 2 are associated with grade and prognosis of human renal carcinoma. APMIS. 2006 May;114(5):329-37. PMID: 16725008 Antioxidant enzymes in renal cell carcinoma. Histol Histopathol. 2006 Feb;21(2):157-65. PMID: 16329040 You might consider ab4867: https://www.abcam.com/index.html?datasheet=4867 I also searched PubMed for nitrotyrosine and lupus given its association with inflammation but only found a paper showing evidence of anti-nitrotyrosine antibodies in the sera of lupus patients. Which lupus tissue(s) might have tyrosine nitration is unclear. Antibodies against nitric oxide damaged poly L-tyrosine and 3-nitrotyrosine levels in systemic lupus erythematosus. J Biochem Mol Biol. 2006 Mar 31;39(2):189-96. PMID: 16584635

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Answer

Thank you for your email. To use ab7225 in IHC, I would recommend titrating the antibody from 1:20 - 1:200, but you will have to optimize based upon your results. If you have any additional questions, please contact us again.

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Answer

Thank you for your enquiry. I'm going to need your order number to tell you precisely how old the stock you received is. To use ab7225 in IHC, I would recommend starting at a dilution of 1:10, but you should try a range in order to optimize the best results.

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Question
Answer

Thank you for your enquiry. The following article is a reference for ab7225: 1) Jessica Ter Steege et al: Nitrotyrosine in plasma of celiac disease patients as detected by a new sandwich Elisa: Free Radiac boil Med 25 (8): 9543-963, 1998. For immunohistology, the dilutions to be used depend on the detection system applied. It is recommended that you test the reagent and determine your own optimal dilution. The typical starting working dilution is 1:10. We would like to warn you that it is very important to keep the following in mind. Do not add sodium azide or any nitric containing preservative since this will affect the antibody! This means for every solution used in the test. Furthermore we would like to advise you to use good positive and negative controls. Is it expectable that the pig liver you tested contains Nitrotyrosine? We hope this information will be helpful to you. If you have further questions please do not hesitate to contact us.

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Answer

This antibody has been found to work well in formalin fixed paraffin embedded tissue and is expected to recognise nitrotyrosine in all species. Heat mediated antigen retrieval will probably be beneficial. Please refer to the Antigen Retrieval protocol, linked on the right hand side of the Abcam homepage.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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