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
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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.
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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.
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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
Preservative: 0.02% Sodium azide
Constituent: HEPES buffered saline -
Concentration information loading...
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Purity
Proprietary Purification -
Purification notes
The antibody was produced in vitro using hybridomas grown in serum-free medium, and then purified by biochemical fractionation. Purity >95% by SDS-PAGE. -
Clonality
Monoclonal -
Clone number
7A12AF6 -
Isotype
IgG2b -
Light chain type
kappa -
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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Isotype control
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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. |
|
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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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
- Nitrotyrosine antibody
Images
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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. -
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.
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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 -
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. -
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 (15)
ab110282 has been referenced in 15 publications.
- Sun X et al. TGF-ß1 attenuates mitochondrial bioenergetics in pulmonary arterial endothelial cells via the disruption of carnitine homeostasis. Redox Biol 36:101593 (2020). PubMed: 32554303
- Haney MJ et al. Genetically modified macrophages accomplish targeted gene delivery to the inflamed brain in transgenic Parkin Q311X(A) mice: importance of administration routes. Sci Rep 10:11818 (2020). PubMed: 32678262
- Nagaoka M et al. A Delphinidin-Enriched Maqui Berry Extract Improves Bone Metabolism and Protects against Bone Loss in Osteopenic Mouse Models. Antioxidants (Basel) 8:N/A (2019). PubMed: 31509995
- Sun F et al. Anti-CD24 Antibody-Nitric Oxide Conjugate Selectively and Potently Suppresses Hepatic Carcinoma. Cancer Res 79:3395-3405 (2019). PubMed: 30918001
- Zheng Y et al. Astragaloside IV enhances taxol chemosensitivity of breast cancer via caveolin-1-targeting oxidant damage. J Cell Physiol 234:4277-4290 (2019). PubMed: 30146689