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

MSCatalog No. MS703

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Overview

Product name

Anti-3-Nitrotyrosine antibody [7A12AF6 ]
See all 3-Nitrotyrosine products (4) ...

Description

Mouse monoclonal [7A12AF6 ] to 3-Nitrotyrosine

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

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

Cross reactivity

Reacts with

Mouse, Rat, Cow, Human

Immunogen

Nitrated KLH (Keyhole Limpet Hemocyanin).

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

Properties

Form

Liquid

Storage instructions

Store at +4°C. Do not freeze.

Storage buffer

Preservative: 0.02% Sodium azide
Constituent: HBS

Concentration

Concentration information loading...

Purity

>95% by SDS-PAGE

Purification notes

The antibody was produced in vitro using hybridomas grown in serum-free medium, and then purified by biochemical fractionation.

Clonality

Monoclonal

Clone number

7A12AF6

Isotype

IgG2b

Light chain type

kappa

Research areas

Metabolism >> Types of disease >> Cancer
Metabolism >> Pathways and Processes >> Redox metabolism >> Oxidative stress
Metabolism >> Pathways and Processes >> Metabolic signaling pathways >> Amino acid metabolism
Cell Biology >> Other Antibodies >> Oxidative Stress
Signal Transduction >> Metabolism >> Amino Acids

  • Flow Cytometry - Nitrotyrosine antibody [7A12AF6 ] (ab110282)Flow Cytometry - Nitrotyrosine antibody [7A12AF6 ] (ab110282) image (enlarge)

  • Immunocytochemistry/ Immunofluorescence - Nitrotyrosine antibody [7A12AF6 ] (ab110282)Immunocytochemistry/ Immunofluorescence - Nitrotyrosine antibody [7A12AF6 ] (ab110282) image (enlarge)

  • Western blot - Nitrotyrosine antibody [7A12AF6 ] (ab110282)Western blot - Nitrotyrosine antibody [7A12AF6 ] (ab110282) image (enlarge)

Applications

Show applications key

Our Abpromise guarantee covers the use of ab110282 in the following tested applications.

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

Target

Relevance

Protein tyrosine nitration results in a post-translational modification that is increasingly receiving attention as an important component of nitric oxide signaling. While multiple nonenzymatic mechanisms are known to be capable of producing nitrated tyrosine residues, most tyrosine nitration events involve catalysis by metalloproteins such as myeloperoxidase, eosinophilperoxidase, myoglobin, the cytochrome P-450s, superoxide dismutase and prostacyclin synthase. Various studies have shown that protein tyrosinenitration is limited to specific proteins and that the process is selective. For example, exposure of human surfactant protein A, SP-A, to oxygen-nitrogen intermediates generated by activated alveolar macrophages resulted in specific nitration of SP-A at tyrosines 164 and 166, while addition of 1.2 mMCO 2 resulted in additional nitration at tyrosine 161. The presence of nitrotyrosine-containing proteins has shown high correlation to disease states such as atherosclerosis, Alzheimer’s disease, Parkinson’s disease and amyotrophic lateral sclerosis.

Alternative names

  • MS703 antibody
  • Nitrotyrosine antibody

Anti-3-Nitrotyrosine antibody [7A12AF6 ] images:

  Flow Cytometry - Nitrotyrosine antibody [7A12AF6 ] (ab110282)

Flow Cytometry - 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.

  Immunocytochemistry/ Immunofluorescence - Nitrotyrosine antibody [7A12AF6 ] (ab110282)

Immunocytochemistry/ Immunofluorescence - 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.

  Western blot - Nitrotyrosine antibody [7A12AF6 ] (ab110282)

Western blot - 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 - Nitrotyrosine antibody [7A12AF6 ] (ab110282)

Western blot - 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 - Nitrotyrosine antibody [7A12AF6 ] (ab110282)

Western blot - 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.

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

ab110282 has not yet been referenced specifically in any publications.

Publishing research using ab110282? Please let us know so that we can cite the reference in this datasheet

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