The monoclonal antibody HM.11 recognizes modified amino acid nitrotyrosine in all different species. Nitrotyrosine is formed in tissues in presence of the active metabolite NO and is a stable end product of nitrosylation of tyrosine. Inflammation is characterized by increased nitric oxide (NO) production. NO reacts rapidly with superoxide to form peroxynitrite. At physiological pH and in the presence of transition metals, peroxynitrite undergoes heterolytic cleavage to form hydroxyl anion and nitronium ion, the latter of which nitrates protein tyrosine residues. The presence of nitrotyrosine has been detected in various inflammatory processes including atherosclerotic plaques, Amyotrophic Lateral Sclerosis (ALS) and Multiple Sclerosis (MS). Thus, the presence of nitrotyrosine on proteins can be used as a marker for peroxynitrite formation in vivo and consequently as a marker of NO-mediated tissue damage. The monoclonal antibody HM.11 recognizes nitrotyrosine, both with the free amino acid as well as with proteins containing nitrotyrosine.

Specifications

Catalog number	HM5002
Product type	Monoclonal antibodies
Quantity	50 µg
Formulation	0.5 ml (100 µg/ml) 0.2 µm filtered sterile biotinylated antibody solution in PBS, containing 0.1% bovine serum albumin.
Conjugate	Biotin
Application	Frozen sections, Immuno assays, Paraffin sections, Western blot
Application Notes	F: also cytospins: acetone fixation 10 min -20 °C; block endogenous peroxidase by 0.3 % H2O2 in PBS (or methanol for intracellular staining); blocking with 10% NGS or 5 % BSA for 30 min. use at assay dependent concentration P:10% formalin fixation; 3% H2O2 to block endogenous peroxidases; Citrate buffer pH 6.0 for 1 min at 100 °C as antigen retrieval treatment; use at assay dependent concentration (1:200/400)- (Ref 1) W: reduced and no-reduced samples; block with 5% BSA or skimmed milk, use at assay dependent concentration
Use	For immuno histology and Western blotting dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:10. Warning: keep sterile, do not add sodium azide or any nitric containing preservative since this will affect the antibody!
Immunogen	Nitrated KLH
Isotype	Mouse IgG2b
Positive Control	W: mouse kidney lysate, mouse optic nerve, retina , spinal cord and brain lysates, rat aorta lysateP: human lung tissue
References	1. ter Steege, JCA et al; Nitrotyrosine in plasma of celiac disease patients as detected by a new sandwich ELISA. Free Radic Biol Med 1998, 25: 953 2. Beckman, JS et al; Extensive nitration of protein tyrosines in human atherosclerosis detected by immunohistochemistry. Biol Chem 1994, 375: 81 3. Kooy, NW et al; Evidence for in vivo peroxynitrite production in human acute lung injury. Am J Respir Crit Care Med 1995, 151: 1250 4. Beckman, JS et al; Apperent hydroxyl radical production by peroxynitrite: Implications of endothelial injury from nitric oxide and superoxide. Proc Natl Acad Sci USA 1990, 87: 1620
Storage and Stability	Product should be stored at 4 °C. Under recommended storage conditions, product is stable for one year.
Precautions	For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result with the use of or derivation of this product.

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