D8075-94B
Clone Type
MonoclonalHost
MouseConjugate
BiotinIsotype
IgG1Clone Number
12F1 (N1664A)Grade
Affinity PurifiedApplications
IC IF IHC IP WBShipping Temp
Blue IceStorage Temp
-20°CMouse Anti-DMPO Nitrone Adduct (Biotin)
5,5-dimethyl-2-(8-octanoic acid)-1-pyrroline N oxide
The formation of free radicals and other highly reactive oxygen species has been implicated in the pathogenesis of many disease states. The ability to identify these species is crucial, and spin trapping has accomplished this goal. DMPO (5,5-dimethyl-1-pyrroline N-oxide) is one of the least toxic to cells and animals, and possesses convenient pharmacokinetics (uptake, distribution, metabolism and excretion) in biological systems. Recent studies have determined that nitric oxide may substantially affect the quantitative determination of DMPO adducts, and therefore extra caution is required when studying generation of these species in the presence of nitric oxide or its radicals. DMPO adducts can be generated with protein and DNA radicals.
Applications
Suitable for use in Western Blot, Immunoprecipitation, Immunohistochemistry, Immunocytochemistry and Immunofluorescence. Other applications not tested.
Recommended Dilutions
Western Blot: 1ug/ml detects the DMPO nitrone adducts of metmyoglobin when loaded at 100ng/lane by colorimetric blot analysis using IgG (HRP) goat anti-mouse as the secondary antibody Optimal dilutions to be determined by the researcher.
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 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Immunogen
5,5-dimethyl-2-(8-octanoic acid)-1-pyrrolone-N-oxide conjugated to Ovalbumin.
Form
Supplied as a liquid in PBS, pH 7.2, 0.09% sodium azide, 50% glycerol. Labeled with Biotin.
Purity
Purified by Protein G affinity chromatography.
Specificity
Recognizes DMPO Nitrone Adduct AT ~90kD. Also recognizes DMPO-octanoic acid, DMPO-protein adducts and DMPO-DNA adducts. Does not react with non-adducted proteins or DNA.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
References
1. Reszka K.J., et al. (2006) Nitric Oxide 15: 133-141, 2. Ramirez D.C., Gomez-Mejiba S.E., and Mason R.P. (2007) Nat Protoc. 2(3): 512-522, 3. Khan N., et al. (2003) Free Radic. Biol. Med 34:1473– 1481, 4. Haseloff R.F., et al. (1997) FEBS Lett 418:73–75, 5. Schaefer C.F., Janzen E.G., West M.S., Poyer J.L., and Kosanke S.D. (1996) Free Radic. Biol. Med 21:427–436, 6. Anzai K., et al. (2003) Arch. Biochem. Biophys 415:251– 256, 7. Free Radic Biol Med. 2009 April 1; 46(7): 853–865. doi: 10.1016/ j.freeradbiomed.2008.12.020.USBio References
No references available