Anti-3-Nitrotyrosine antibody [7A12AF6] (ab110282)
- Datasheet
- Specific References (6)
- Protocols
Overview
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Product nameAnti-3-Nitrotyrosine antibody [7A12AF6]
See all 3-Nitrotyrosine primary antibodies -
DescriptionMouse monoclonal [7A12AF6] to 3-Nitrotyrosine
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Host speciesMouse
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Specificityab110282 was developed to recognize only protein-bound nitrotyrosine and so is a sensitive tool for measuring protein-specific modifications from oxidative stress.
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Tested applicationsSuitable for: ICC/IF, Flow Cyt, WB, IP, In-Cell ELISAmore details
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Species reactivityReacts with: Mouse, Rat, Cow, Human
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Immunogen
Nitrated KLH (Keyhole Limpet Hemocyanin).
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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
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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.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C. Do Not Freeze.
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Storage bufferPreservative: 0.02% Sodium azide
Constituent: HEPES buffered saline -
Concentration information loading... -
Purity>95% by SDS-PAGE
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Purification notesThe antibody was produced in vitro using hybridomas grown in serum-free medium, and then purified by biochemical fractionation.
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ClonalityMonoclonal
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Clone number7A12AF6
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IsotypeIgG2b
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Light chain typekappa
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Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Related Products
Applications
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.
| Application | Abreviews | Notes |
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| ICC/IF | Use a concentration of 1 µg/ml. | |
| Flow Cyt | Use a concentration of 1 µg/ml. ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
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| WB | Use a concentration of 1 µg/ml. | |
| IP | Use at an assay dependent concentration. | |
| In-Cell ELISA | Use a concentration of 4 µg/ml. (0.4 µg/well). |
Target
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RelevanceProtein 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.
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Alternative names
- Nitrotyrosine antibody
Images
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![Immunocytochemistry/ Immunofluorescence - Anti-3-Nitrotyrosine antibody [7A12AF6] (ab110282)](https://www.abcam.com/ps/products/110/ab110282/Images/Nitrotyrosine-Primary-antibodies-ab110282-7.jpg)
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)](https://www.abcam.com/ps/products/110/ab110282/Images/Nitrotyrosine-Primary-antibodies-ab110282-6.jpg)
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)](https://www.abcam.com/ps/products/110/ab110282/Images/Nitrotyrosine-Primary-antibodies-ab110282-9.jpg)
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)](https://www.abcam.com/ps/products/110/ab110282/Images/Nitrotyrosine-Primary-antibodies-ab110282-10.jpg)
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)](https://www.abcam.com/ps/products/110/ab110282/Images/Nitrotyrosine-Primary-antibodies-ab110282-11.jpg)
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.
Protocols
References
This product has been referenced in:
- Kehm R et al. Age-related oxidative changes in pancreatic islets are predominantly located in the vascular system. Redox Biol 15:387-393 (2018). Read more (PubMed: 29331666) »
- Moghimi E et al. Dietary Vitamin D3 Restriction Exacerbates Disease Pathophysiology in the Spinal Cord of the G93A Mouse Model of Amyotrophic Lateral Sclerosis. PLoS One 10:e0126355 (2015). WB ; Mouse . Read more (PubMed: 26020962) »
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"