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Anti-3-Nitrotyrosine antibody [7A12AF6] (ab110282)

Key features and details

  • Mouse monoclonal [7A12AF6] to 3-Nitrotyrosine
  • Suitable for: ICC/IF, Flow Cyt, WB, IP, In-Cell ELISA
  • Reacts with: Mouse, Rat, Cow, Human
  • Isotype: IgG2b

Overview

  • Product name

    Anti-3-Nitrotyrosine antibody [7A12AF6]
    See all 3-Nitrotyrosine primary antibodies

  • Description

    Mouse monoclonal [7A12AF6] to 3-Nitrotyrosine

  • Host species

    Mouse

  • Specificity

    ab110282 was developed to recognize only protein-bound nitrotyrosine and so is a sensitive tool for measuring protein-specific modifications from oxidative stress.

  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, WB, IP, In-Cell ELISAmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Cow, Human

  • Immunogen

    Chemical/ Small Molecule. This information is considered to be commercially sensitive.

  • Positive control

    • Flow Cyt: HL-60 cells treated with 2 mM peroxynitrite ICC/IF: HeLa cells and Human fibroblast cells treated with 1 mM peroxynitrite WB: nitrated Bovine heart mitochondria; nitrated tyrosine

  • General notes

    This antibody clone is manufactured by Abcam.

    Product was previously marketed under the MitoSciences sub-brand.

    Anti-3-Nitrotyrosine antibody (Alexa Fluor® 488) [7A12AF6] (ab157402)

    Anti-3-Nitrotyrosine antibody (HRP) [7A12AF6] (ab198491)

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

    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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-3-Nitrotyrosine antibody [7A12AF6] (ab110282)
    Immunocytochemistry/ Immunofluorescence - Anti-3-Nitrotyrosine antibody [7A12AF6] (ab110282)
    Immunocytochemistry image of ab110282 stained Human HeLa cells (A) and fibroblast cells (B, C).
    Cells grown on slides were paraformaldehyde fixed (4%, 20 min) and Triton X-100 permeabilized (0.1%, 15 min). Slides were treated with/without 1 mM peroxynitrite to modify exposed tyrosines to 3-nitrotyrosine. Slides were blocked and incubated with tab110282 at 2 µg/ml overnight at 4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Mouse IgG (H+L) used at a 1/1000 dilution for 1 hour. 10% Goat serum was used as the blocking agent for all blocking steps. For reference, the mitochondria (red) were identified by HSP60 / Alexa Fluor® 594 and DAPI was used to stain the cell nuclei (blue).
    HeLa cells (A) and fibroblast cells (B) show surface modification of tyrosine to 3-nitrotyrosine after exposure to peroxynitrite. While (C) unexposed fibroblast cells show no modification.
  • Flow Cytometry - Anti-3-Nitrotyrosine antibody [7A12AF6] (ab110282)
    Flow Cytometry - Anti-3-Nitrotyrosine antibody [7A12AF6] (ab110282)
    HL-60 cells were stained with 1 µg/ml ab110282 following treatment with 2 mM peroxynitrite (blue) or vehicle control (red). No primary antibody control is shown in black. Peroxynitrite modifies tyrosine residues to 3-nitrotyrosine.
  • Western blot - Anti-3-Nitrotyrosine antibody [7A12AF6] (ab110282)
    Western blot - Anti-3-Nitrotyrosine antibody [7A12AF6] (ab110282)
    All lanes : Anti-3-Nitrotyrosine antibody [7A12AF6] (ab110282) at 1 µg/ml

    Lane 1 : Bovine heart mitochondria
    Lane 2 : Bovine heart mitochondria - nitrated
    Lane 3 : BSA
    Lane 4 : BSA - nitrated
  • Western blot - Anti-3-Nitrotyrosine antibody [7A12AF6] (ab110282)
    Western blot - Anti-3-Nitrotyrosine antibody [7A12AF6] (ab110282)
    All lanes : Anti-3-Nitrotyrosine antibody [7A12AF6] (ab110282) at 1 µg/ml

    Lane 1 : Bovine heart mitochondria
    Lane 2 : Bovine heart mitochondria - nitrated
    Lane 3 : BSA
    Lane 4 : BSA - nitrated


    Prior to runnning the samples, the membrane was treated with sodium dithionite to reduce nitrotyrosine to aminotyrosine.
  • Western blot - Anti-3-Nitrotyrosine antibody [7A12AF6] (ab110282)
    Western blot - Anti-3-Nitrotyrosine antibody [7A12AF6] (ab110282)
    All lanes : Anti-3-Nitrotyrosine antibody [7A12AF6] (ab110282) at 1 µg/ml

    Lane 1 : Bovine heart mitochondria
    Lane 2 : Bovine heart mitochondria - nitrated
    Lane 3 : BSA
    Lane 4 : BSA - nitrated


    In this experiment, the antibody was first blocked with free nitrotyrosine before being used to blot the membrane.
    The figure shows that ab110282's binding capacity was not inhibited by the free nitrotyrosine, and so only binds to the protein-bound form.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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