Technical Data
N2700-06
Nitrotyrosine (nTyr)
Description:
Nitrotyrosine has been identified as an indicator of cell damage and inflammation, as well as of the production of NO. It is believed that measuring the concentration of nitrotyrosine will serve as a marker for damage caused by NO in the cell. Previous studies have shown that nitrotyrosine has been found in inflammatory conditions such as atherosclerotic plaques and rheumatoid arthritis, among others.

Applications:
Suitable for use in Western Blot, Immunohistochemistry and Immunocytochemistry. Other applications not tested.

Recommended Dilutions:
Western Blot: 0.5-2ug/ml detects nitrotyrosine immunoblot controls N2705 at 16kD, 66kD and 215kD, corresponding to nitrated superoxide dismutase, nitrated BSA, and nitrated rabbit muscle myosin, respectively. An ~32kD band might be seen representing a dimer of the 16kD protein. Nitrotyrosine immunoblot controls were resolved by electrophoresis, transferred to nitrocellulose and probed with N2700-06 (2ug/ml). Proteins were visualized using an IgG (HRP) goat anti-rabbit and a chemiluminescence detection system.
Immunohistochemistry: 5ug/ml detects nitrated proteins in formalin-fixed paraffin embedded lateral ischemic rat brain tissue. Has been reported to detect nitrotyrosine-containing proteins in acutely injured human lung tissue.
Immunocytochemistry: 5-10ug/ml detects nitrated proteins from human A431 cells pretreated 5 minutes with 5-10mM peroxynitrite.
Optimal dilutions to be determined by the researcher.

Positive Controls:
Immunohistochemistry: Human Alzheimer disease brains sections (Methacarn fixed) and mouse liver
Western Blot: Nitrated-BSA

Storage and Stability:
May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
TypeIsotypeCloneGrade
PabIgGAffinity Purified
SizeStorageShippingSourceHost
100ug-20CBlue IceRabbit
Concentration:
~1mg/ml
Immunogen:
Nitrated Keyhole Limpet Hemocyanin. Epitope: Nitrated Tyr
Purity:
Purified by immunoaffinity chromatography.
Form
Supplied as a liquid in 0.1M Tris-Glycine, pH 7.4, 0.15M sodium chloride, 10mg/ml BSA, 0.05% sodium azide.
Specificity:
Recognizes nitrated proteins from all species.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
US Biological application reference: Jaubert, A.-M. et al., (2012) PLoS ONE 7(7): e40650. doi:10.1371/journal.pone.0040650. 1. Beckman, J.S., et al., Biol. Chem. Hoppe-Seyler 375:8188 (1994). 2. Ohshima, H., et al., Fund. Chem. Tox. 28:647652 (1990). 3. Ischiropoulos, H., et al., Arch. Biochem. Biophys. 298:431437 (1992). 4. Beckman, J.S., et al., Nature 364:584 (1993). 5. Kooy, N.W., et al., Am. J. Respir. Crit. Care Med. 151:12501254 (1995). 6. Rameau, G.A., et al. (2004). J Biol Chem. 279: 14307-14. 7. Murray, J., et al. (2003). J Biol Chem. 278: 37223-30. 8. Aboul-Enein, Fahmy, et al. (2006). Acta Neuropathol. 111: 539-47. 9. Gaut, J.P., et al. (2002). J Clin Invest. 109: 1311-9. 10. Aboul-Enein, Fahmy, et al. (2004). J. Neuropathol. Exp. Neurol. 63: 1284-96.