The cellular production of highly reactive nitrogen species derived from nitric oxide, such as peroxynitrite, nitrogen dioxide and nitryl chloride, leads to the nitration of tyrosine resides in tissue proteins. The extent of protein nitrotyrosine formation provides an index of the production of reactive nitrogen species and potential cell damage over a period of time. Nitrotyrosine can be measured by amino-acid analysis of protein hydrolysates and detected, estimated semi-quantitatively and located in cells and tissues by immunocytochemical techniques using antibodies directed against the nitrotyrosine hapten.

Nitrated KLH.

Detect Nitrotyrosine using this Anti-Nitrotyrosine Antibody validated for use in IH(P) & WB.

Research Category
Neuroscience

Research Sub Category
Oxidative Stress

Western Blot:
1:100-1:1,000 dilution of a previous lot was used sucessfully in western blot when tested against nitrated BSA and nitrated ovalbumin.

Immunohistochemistry:
1:50-1:100 on paraffin embedded, 10% neutral buffer formalin fixed mouse liver (animal treated with Zymosan) and normal human cerebellum tissues. The tissues must be pretreated with heat induced epitope retrieval (HIER). Incubation with the AB5411 was for 10 minutes and the reaction was developed with the Chemicon IHCSelect, HRP-DAB kit (Catalog number DAB150).

Optimal working dilutions must be determined by end user.

Reactivity with other species has not been determined.

Recognizes nitrated tyrosine residues.

Format: Purified

Protein A purified

Purified rabbit polyclonal in buffer containing 0.1% sodium azide.

Stable for 1 year at 2-8ºC from date of receipt.

Control
POSITIVE CONTROL: Tyrosine residues can be nitrated directly in situ on tissue sections using peroxynitrite (catalog # 20-107) to create a positive control. After the tissue section is deparaffinized, the slide is covered with a drop of PBS, pH 7.4 into which 10 microliters of peroxynitrite solution are mixed. Care should be taken that the solution maintains a pH of 7.4.

Immunohistochemistry(paraffin) Analysis:
Nitrotyrosine (AB5411) staining of Mouse Brain (Zymosan), tissue pretreated with citrate buffer, pH 6.0. Polyclonal antibody was diluted to 1:500, using IHC-Select Detection with HRP-DAB. Immunoreactivity is seen in here in hippocampus.
Optimal Staining of Nitrotyrosine Monoclonal: Mouse Brain Treated With Zymosan.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Replaces: AB5386

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