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Specifications

Clone Host Grade Applications Monoclonal Mouse Affinity Purified E B IH

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.
Catalog #	N2700-03
Applications	Suitable for use in ELISA, Western Blot and Immunohistochemistry. Other applications have not been tested.
Recommended Dilutions	Western Blot: 1:50 for reduced and no-reduced samples. Block with 5% BSA or skimmed milk.
	Immunohistochemistry (Frozen, cytospins): 1:50 with acetone fixation for 10 minutes at -20 °C. Block endogenous peroxidase with 0.3% H2O2 in PBS (or methanol for intracellular staining). Block with 10% NGS or 5% BSA for 30 minutes.
	Immunohistochemistry (Paraffin): 1:200-1:400 with 10% formalin fixation, Use 3% H2O2 to block endogenous peroxidases. Use citrate buffer pH 6.0 for 1 minute at 100°C as antigen retrieval treatment.
	Optimal dilutions to be determined by the researcher.
Western Blot Controls	Mouse kidney lysate, mouse optic nerve, retina , spinal cord and brain lysates, rat aorta lysate
Storage and Stability	May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
CAS Number	n/a
Clone Type	Monoclonal
Isotype	IgG2b
Clone No	6A610
Host	Mouse
Concentration	~0.1mg/ml
Form	Supplied as a liquid in PBS, 0.1% BSA, 0.02% sodium azide.
Purity	Purified
Immunogen	Nitrated KLH
Specificity	Recognizes nitrotyrosine, both with the free amino acid as well as with proteins containing nitrotyrosine. Does not crossreact with Phosphotyrosine or Chlorotyrosine.
Important Note	This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.