Nitrotyrosine, mAb HM.11

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Description
Application

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

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

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 1:50.

Positive Control

W: mouse kidney lysate, mouse optic nerve, retina , spinal cord and brain lysates, rat aorta lysate P: human lung tissue

Specifications
Downloads

Product type

Monoclonal antibodies

Amount

100 µg, 20 µg, 500 µg

Formulation

Quantities >500 µg will be send without preservative and carrier free. Quantities <500 µg will be send with preservative and carrier. exact formulation can be found on the datasheet.

Immunogen

Nitrated KLH

Isotype

Mouse IgG2b

Species

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.

Disease

Autoimmunity, Cardiology and metabolism

Protocol