DMPO

Free radicals are highly reactive and generally short-lived species. Hence, most studies of free radicals have been restricted to analysis of the end products formed by their interaction with cellular biomolecules. One technology that has permitted direct study of free radicals is electron spin resonance (ESR). For very short lived radicals, spin traps have been empolyed to generate longer lived radicals that are more amenable to ESR analysis. The most widely used of these spin traps is 5,5-dimethyl-1-pyrroline-N-oxide (DMPO). The reaction of DMPO with a protein radical gives rise to a nitroxide radical which can be disproportionate to hydrozylamine and nitrone adducts. Of these species, only the nitrone adduct can be considered stable. DMPO antibodies will specifically react with radicals in protein.\n\nApplications: \nSuitable for use in ELISA, Western Blot and Immunocytochemistry. Other applications not tested.\n\nRecommended Dilutions:\nELISA: 1:5000. Reactive against DMPO-protein adducts in dilutions of 1:100-10,000 followed by reaction with alkaline phosphatase conjugated, affinity purified goat anti-rabbit IgG.\nWestern Blot: 1:5000\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees 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.

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SPECIFICATIONS

Catalog Number

D8075-94

Size

50ul

Applications

ELISA, ICC, WB

Hosts

Rabbit

Form

Supplkied as a liquid. No stabilizers or preservatives added.

P Type

Pab

Purity

Ascites

Isotype

IgG1

References

11. Immunological identification of the heart myoglobin radical formed by hydrogen peroxide: C.D. Detweiler, et al.; Free Radic. Biol. Med. 33, 364 (2002). 2. Immunochemical detection of hemoglobin-derived radicals formed by reaction with hydrogen peroxide: involvement of a protein-tyrosyl radical: D.C. Ramirez, et al.; Free Radic. Biol. Med. 34, 830 (2003). 3. UVA-ketoprofen-induced hemoglobin radicals detected by immuno-spin trapping: Y.Y. He, et al.; Photochem. Photobiol. 77, 585 (2003). 4. Reaction of human hemoglobin with peroxynitrite. Isomerization to nitrate and secondary formation of protein radicals: N. Romero, et al.; J. Biol. Chem. 278, 44049 (2003). 5. Identification of free radicals on hemoglobin from its self-peroxidation using mass spectrometry and immuno-spin trapping: observation of a histidinyl radical: L. Deterding; J. Biol. Chem. 279, 11600 (2004). 6. Using anti-5,5-dimethyl-1-pyrroline N-oxide (anti-DMPO) to detect protein radicals in time and space with immuno-spin trapping: R.P. Mason; Free Radic. Biol. Med. 36, 1214 (2004). 7. Protein radical formation during lactoperoxidase-mediated oxidation of the suicide substrate glutathione: immunochemical detection of a lactoperoxidase radical-derived 5,5-dimethyl-1-pyrroline N-oxide nitrone adduct: Q. Guo, et al.; J. Biol. Chem. 279, 13272 (2004). 8. Involvement of protein radical, protein aggregation, and effects on NO metabolism in the hypochlorite-mediated oxidation of mitochondrial cytochrome c: Y.R. Chen, et al.; Free Radic. Biol. Med. 37, 1591 (2004). 9. Novel identification of a sulfur-centered, radical-derived 5,5-dimethyl-1-pyrroline N-oxide nitrone adduct formed from the oxidation of DTT by LC/ELISA, LC/electrospray ionization-MS, and LC/tandem MS: Q. Guo, et al.; Chem. Res. Toxicol. 17, 1481 (2004). 10. Mechanism of hydrogen peroxide-induced Cu,Zn-superoxide dismutase-centered radical formation as explored by immuno-spin trapping: the role of copper- and carbonate radical anion-mediated oxidations: D.C. Ramirez, et al.; Free Radic. Biol. Med. 38, 201 (2005)

Additional Info

Recognizes DMPO, DMPO-octanoic acid and DMPO-protein adducts. Does not cross react with non-adducted proteins.