Biotin Anti-Nitro tyrosine antibody [HM.11] (ab7225)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Biotin Anti-Nitro tyrosine antibody [HM.11] (ab7225)

    Key features and details

    • Biotin Mouse monoclonal [HM.11] to Nitro tyrosine
    • Suitable for: IHC-P
    • Reacts with: Human
    • Conjugation: Biotin
    • Isotype: IgG2b

    Overview

    • Product name

      Biotin Anti-Nitro tyrosine antibody [HM.11]
      See all Nitro tyrosine primary antibodies

    • Description

      Biotin Mouse monoclonal [HM.11] to Nitro tyrosine

    • 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: IHC-Pmore details

    • Species reactivity

      Reacts with: Human
      Predicted to work with: Mouse, Rat

    • General notes

      Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

      Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

      We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

      In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

      We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

      Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

      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, as well as customer reviews and Q&As.

    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
    IHC-P 1/200 - 1/400.

    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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Biotin Anti-Nitro tyrosine antibody [HM.11] (ab7225)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Biotin Anti-Nitro tyrosine antibody [HM.11] (ab7225)

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

    References (2)

    Publishing research using ab7225? Please let us know so that we can cite the reference in this datasheet.

    ab7225 has been referenced in 2 publications.

    • 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 . 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 . PubMed: 15878361

    Customer reviews and Q&As

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    1-6 of 6 Abreviews or Q&A

    Question

    I would like to find out if you have information on dilution for westerns. At least to get a starting point or range. thanks

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    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

    What tissue should I use for a positive IHC control?

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    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/Kidney-tumor-human-renal-cell-carcinoma-tissue-slides-ab4867.html 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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    Question

    I want to know the consentration of mouse monoclonal to Nitrotyrosine (ab7225) in the immunohistochemistry immediately. Can you reply my email immediately?

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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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    Question

    ANTIBODY CODE ab7225 BATCH NUMBER 4915M13 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM We have this antibody in our possession and would like to know a) what titer to try on paraffin embedded mouse lung, and b) how old is this particular lot.

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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

    I have answered your questions. Many thanks 1. Please describe the problem (high background, no staining etc). no staining 2. On what material are you testing the antibody in IHC? * Species? - pig * Cell line?- liver * Tissue? - liver 3. How did you fix the samples? Acetone 4. Did you apply antigen retrieval step? Other - none 5. How did you block the unspecific binding sites? porcine serum 6. Primary antibody * Specification (in which species was it raised against)? Mouse * At what dilution(s) have you tested this antibody? 1:50 * Incubation time, wash steps (multiple short washes are more effective than fewer longer wash steps)? Over night, Wash with TBS 7. Secondary antibody * What secondary antibody are you using? anti-mouse * Specification (in which species was it raised against)? Rabbit * At what dilution(s) have you tested this antibody? 1: 50 * Incubation, wash steps? TBS * Do you know whether the problems you are experiencing come from the secondary? no * What detection method are you using? APAAP 8. Background staining * Please provide an image of your staining 9. Which detection system did you use? Fast red TR 10. Did you apply positive and negative controls along with the samples? Please specify. None for nitrotyrosine 11. Optimization attempts * How many times have you tried the IHC? twice * Do you obtain the same results every time? Yes * What steps have you altered? Incubate with secondary and APAAP complex twice

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    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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    Question

    Does this antibody work well in paraffin? Does it work in rats and mice? What are suggested fixatives and pretreatments?

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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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