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  1. Link

    nitro-tyrosine-antibody-hm11-ab7048.pdf

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Neuroscience Neurotransmission Nitric Oxide Nitrated Species
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Anti-Nitro tyrosine antibody [HM.11] (ab7048)

  • Datasheet
  • SDS
Reviews (13)Q&A (21)References (64)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nitro tyrosine antibody [HM.11] (ab7048)

    Key features and details

    • Mouse monoclonal [HM.11] to Nitro tyrosine
    • Suitable for: IHC-Fr, IHC-P
    • Reacts with: Species independent
    • Isotype: IgG2b

    Conjugates logo Related conjugates and formulations

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    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: IHC-Fr, IHC-Pmore details
    • Species reactivity

      Reacts with: Species independent
    • 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)
    • General notes

      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

    Properties

    • Form

      Liquid
    • Storage instructions

      Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
    • Storage buffer

      Constituents: PBS, 0.1% BSA

      WARNING!!: Do not add sodium azide or nitric containing perservative as it will damage the antibody.
    • Concentration information loading...
    • Purity

      Protein G purified
    • Clonality

      Monoclonal
    • Clone number

      HM.11
    • Isotype

      IgG2b
    • Research areas

      • Neuroscience
      • Neurotransmission
      • Nitric Oxide
      • Nitrated Species
      • Cardiovascular
      • Atherosclerosis
      • Thrombosis
      • Platelets

    Associated products

    • Alternative Versions

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

      • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
      • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Isotype control

      • Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (ab170192)

    Applications

    The Abpromise guarantee

    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
    IHC-Fr
    Use at an assay dependent concentration.

    Acetone fixation 10 min -20 degree 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.

    IHC-P (3)
    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
    Notes
    IHC-Fr
    Use at an assay dependent concentration.

    Acetone fixation 10 min -20 degree 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.

    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

    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) - 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.

    Protocols

    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (64)

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

    ab7048 has been referenced in 64 publications.

    • Liou GG  et al. N-Acetyl Cysteine Overdose Inducing Hepatic Steatosis and Systemic Inflammation in Both Propacetamol-Induced Hepatotoxic and Normal Mice. Antioxidants (Basel) 10:N/A (2021). PubMed: 33809388
    • Clayton ZS  et al. Apigenin restores endothelial function by ameliorating oxidative stress, reverses aortic stiffening, and mitigates vascular inflammation with aging. Am J Physiol Heart Circ Physiol 321:H185-H196 (2021). PubMed: 34114892
    • Qin X  et al. Chronic overload of concentration-dependent iron exerts different effects on ovarian function in C57BL/6J mice†. Biol Reprod 104:1347-1359 (2021). PubMed: 33693648
    • Ramos H  et al. Beneficial Effects of Glucagon-Like Peptide-1 (GLP-1) in Diabetes-Induced Retinal Abnormalities: Involvement of Oxidative Stress. Antioxidants (Basel) 9:N/A (2020). PubMed: 32927585
    • Gioscia-Ryan RA  et al. Late-life voluntary wheel running reverses age-related aortic stiffness in mice: a translational model for studying mechanisms of exercise-mediated arterial de-stiffening. Geroscience N/A:N/A (2020). PubMed: 32529594
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    11-20 of 34 Abreviews or Q&A

    Western blot abreview for Anti-Nitro tyrosine antibody [HM.11]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (liver)
    Loading amount
    50 µg
    Specification
    liver
    Treatment
    warm ischemia/reperfusion
    Gel Running Conditions
    Reduced Denaturing (12)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    Submitted May 08 2012

    Western blot abreview for Anti-Nitro tyrosine antibody [HM.11]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (aorta)
    Loading amount
    20 µg
    Specification
    aorta
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 01 2011

    Western blot abreview for Anti-Nitro tyrosine antibody [HM.11]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Tissue lysate - whole (Human Penile Tissue)
    Loading amount
    60 µg
    Specification
    Human Penile Tissue
    Gel Running Conditions
    Reduced Denaturing (4-20% gel)
    Blocking step
    Starting Block T20 (PBS) as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
    Read More

    Miss. Gwen Lagoda

    Verified customer

    Submitted Oct 19 2011

    Western blot abreview for Anti-Nitro tyrosine antibody [HM.11]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (liver)
    Loading amount
    20 µg
    Specification
    liver
    Treatment
    Acetaminophen for 3 hrs
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 06 2011

    Western blot abreview for Anti-Nitro tyrosine antibody [HM.11]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Penile Tissue)
    Loading amount
    30 µg
    Specification
    Penile Tissue
    Gel Running Conditions
    Reduced Denaturing (4-20% mini gel)
    Blocking step
    Starting Block T20 (PBS) as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
    Read More

    Miss. Gwen Lagoda

    Verified customer

    Submitted Dec 08 2008

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

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Liver)
    Specification
    Liver
    Fixative
    Paraformaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer 1X
    Permeabilization
    No
    Blocking step
    Peroxidase Blocking Reagent as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
    Read More

    Miss. Gwen Lagoda

    Verified customer

    Submitted Jan 23 2008

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (Penile Tissue)
    Specification
    Penile Tissue
    Fixative
    Paraformaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer 1X
    Permeabilization
    No
    Blocking step
    Peroxidase Blocking Reagent as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
    Read More

    Miss. Gwen Lagoda

    Verified customer

    Submitted Dec 10 2007

    Western blot abreview for Anti-Nitro tyrosine antibody [HM.11]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Lung)
    Loading amount
    10 µg
    Specification
    Lung
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 23 2007

    Question

    Hello! I am sorry that I give the response late. As I told you in the last e-mail, I took 4 weeks holiday last month. I came back to work last week and made a few runs for the nitrotyronsine (ab7048) staining following your recommendation. I am sorry to tell that I did not get any signal from the antibody even the triton was added to the blocking and dilution buffer (0,3%v/v). And I also try some new prepared and less fixed (16 hrs) samples (70% liver partial hepatectomy). Unfortunately, there is still no signal. Based on the literature, the 70% liver partial hepatectomy sample should be positive with this clone of antibody, although this paper used the antibody from zymed. Therefore, I came back to you again for further help. Thanks a lot for your kindly help in advance.

    Read More

    Abcam community

    Verified customer

    Asked on Mar 10 2008

    Answer

    I'm sorry to hear you are still having problems with ab7048. I would be happy to offer you a replacement vial or refund if the antibody was purchased less than 120 days before you first contacted us on the 21st Jan. If you can please let me know your order details and what you would prefer I will arrange this for you.

    Read More

    Abcam Scientific Support

    Answered on Mar 10 2008

    Question

    BATCH NUMBER 408786 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM No staining SAMPLE 3 kinds of rat livers (Lewis) 1) 1 week after transplantation, 2)1 week and 4 weeks after 70% partial hepatectomy 3) 48 hours after right hepatic vein ligation Rat lung (lewis) with vasculitis PRIMARY ANTIBODY Nitrotyrosine (clone HM11) ( Cat: ab7048, Lot: 408786) 1 to 20, 1 to 30, 1 to 50 dilution No wash after the blocking, directly add the diluted primary antibody to the samples. Incubate at RT for 1 hr.Wash with wash buffer 3 x 3 min thereafter. DETECTION METHOD Fast Red kit (Dako, ref: k0597, lot: 00004136 from 08-2007) POSITIVE AND NEGATIVE CONTROLS USED no positive control is availabe. The negative control was negative and clean. ANTIBODY STORAGE CONDITIONS It arrived on 8th, Jan, 2008 at 4 °C and was aliquoted by me in steril eppendorf tubes storing at -20°C. FIXATION OF SAMPLE Paraformaldehyde 48 hrs or For frozen section Cold Acetone 10mins plus 90% enthanl 5 mins ANTIGEN RETRIEVAL EDTA buffer pH 8,0 Citrate EDTA buffer pH 6,2 95°C for 10 mins PERMEABILIZATION STEP no BLOCKING CONDITIONS Protein block (DakoCytomation, Code#X0909, Lot#036049) 1 hour, room temperature SECONDARY ANTIBODY Power Vision mono AP-anti-mouse IgG (Immunologic, Cat# DPVM-110AP, Lot# 50212, from 2007)Incubate at RT for 30 mins.Wash with wash buffer 3 x 3 min thereafter. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? many HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? sample selection and antigen retrieval buffer

    Read More

    Abcam community

    Verified customer

    Asked on Jan 22 2008

    Answer

    Thank you for your enquiry. I am sorry to hear you are having a problem with ab7048. I think the protocol you have been using for paraffin sections is very good, and have only a few recommendations to make, which I hope will help resolve the lack of staining: -I would recommend to incubate the primary antibody overnight to maximise the binding time -the PFA fixation may be too strong, typically tissue is fixed for 24 hours. Would you have access to tissue less fixed, to see if the epitope is modified from the fixation? -I would recommend adding some triton to the blocking and dilution buffer (0.3%v/v) to enable the antibodies to penetrate into the tissue more easily For frozen sections, can I please check these do not have an antigen retrieval step? I would recommend trying 4% PFA fixation (10mins) and following the same tips as above. I hope these recommendations help; if you still do not obtain a successful staining, please let me know and I would be happy to offer you a replacement vial or refund.

    Read More

    Abcam Scientific Support

    Answered on Jan 22 2008

    11-20 of 34 Abreviews or Q&A

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