DMPO, pAb ALX-210-530-R050 50 µl
Detection of protein-derived radicals in isolated proteins has Protein + Cu(II) + H2O2
typically required the utilization of direct electron spin resonance
(ESR) and/or spin trapping ESR. ESR can be applied both to
understand the mechanisms of protein radical formation and
for structural identiﬁcation of the amino acid-derived radical in- Protein-centered Radicals
volved. However, ESR measurements require signiﬁcant levels (t 1/2 =µsec-sec)
of protein and of its derived radicals (greater than micromolar) + DMPO
and rely on rather specialized and costly equipment. Moreover,
detection of protein radicals by ESR becomes a difﬁcult task in DMPO-protein Radical Adducts
cells and tissues. (t 1/2 =sec-min)
Immuno-spin trapping is based on the fact that certain amino - e-
acid-derived radicals (most notably, but not exclusively, tyrosyl
radicals) can react with the spin trap 5,5-dimethyl-1-pyrroline-N- DMPO-protein Radical-derived
Nitrone Adducts Immuno-spin Trapping
oxide (DMPO) to form a protein-DMPO nitroxide radical adduct, (t 1/2 =days-months)
which can then be oxidized by one electron to the corresponding
ESR-silent protein-DMPO nitrone adduct. Indeed, DMPO spin FIGURE: Schematic overview of approaches to detect protein radicals.
adducts are not stable, have a half-life on the order of minutes,
and tend to decay under oxidizing conditions to the rather stable
DMPO nitrone. Rabbit polyclonal anti-DMPO nitrone antiserum
P. Mason: Immuno-spin trapping of DNA radicals; Nat. Methods
(Prod. No. ALX-210-530) has been developed and validated in
3, 127 (2006)), which broadens even more the scope of applica-
Mason’s laboratory (C. D. Detweiler, L. J. Deterding, K. B. Tomer,
tions of this breakthrough technique in free radical biology and
C. F. Chignell, D. Germolec, R. P. Mason: Immunological identiﬁ-
cation of the heart myoglobin radical formed by hydrogen perox-
ide; Free Radic. Biol. Med. 33, 364 (2002)) and a series of studies Adapted from: Immuno-spin trapping: A breakthrough for the
have shown its utility to demonstrate protein radical formation in sensitive detection of protein-derived radicals, a commentary on
different proteins exposed to a variety of oxidants. “Protein radical formation on thyroid peroxidase during turnover”;
In summary, immuno-spin trapping is proving to be a potent, R. Radi; Free Radic. Biol. Med. 41, 416 (2006).
sensitive, and accessible method to detect low levels (e.g. great-
er than nanomolar) of protein-derived radicals produced in vitro
and potentially, and yet to be established, in vivo. Moreover, it Selected Review Article
has just been established that anti-DMPO nitrone antibodies can 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
be utilized for the detection of DNA-derived radicals in the form (2004)
of DNA-DMPO nitrone adducts (D. C. Ramirez, S. E. Mejiba, R.
International Edition 1-Nov-2009 www.enzolifesciences.com
ALX-210-530-R050 50 µl
From rabbit. IMMUNOGEN: DMPO-octanoic acid conjugated to
ovalbumin. SPECIFICITY: Recognizes DMPO (Prod. No. ALX-
430-090), DMPO-octanoic acid and DMPO-protein adducts.
Does not cross react with non-adducted proteins. APPLICA-
TION: ELISA, IHC, WB.
Widely Published Switzerland & Rest of Europe
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munochemical detection of hemoglobin-derived radicals formed by reaction with
hydrogen peroxide: involvement of a protein-tyrosyl radical: D. C. Ramirez, et al.; CH-4415 Lausen / Switzerland
Free Radic. Biol. Med. 34, 830 (2003) Identification of free radicals on hemo- Tel. + 41/0 61 926 89 89
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(2004) Protein radical formation during lactoperoxidase-mediated oxidation of email@example.com
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) Mechanism of hydrogen peroxide-induced Cu,Zn-
superoxide dismutase-centered radical formation as explored by immuno-spin North/South America
trapping: the role of copper- and carbonate radical anion-mediated oxidations:
D. C. Ramirez, et al.; Free Radic. Biol. Med. 38, 201 (2005) Identification of the ENZO LIFE SCIENCES INTERNATIONAL, INC.
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weiler, et al.; Free Radic. Biol. Med. 38, 969 (2005) Immuno-spin trapping: detec-
tion of protein-centered radicals: D. C. Ramirez, et al.; Curr. Prot. Toxicol. 2, 17.7.1 FIGURE: Confocal microscopy images of the colocalization of Plymouth Meeting, PA 19462-1202
(2005) Superoxide generation from mitochondrial NADH dehydrogenase induces catalase (red stain) and protein-DMPO adducts (green stain) USA
self-inactivation with specific protein radical formation: Y. R. Chen, et al.; J. Biol. obtained by treating mouse hepatocytes (2.5x106 cells/ml) with
Chem. 280, 37339 (2005) Immunochemical detection of nitric oxide and nitrogen HOCl. Tel. 1-800-942-0430 / (610) 941-0430
dioxide trapping of the tyrosyl radical and the resulting nitrotyrosine in sperm whale Fax (610) 941-9252
myoglobin: K. Nakai and R. P. Mason; Free Radic. Biol. Med. 39, 1050 (2005) Picture courtesy of Dr. M. G. Bonini, National Institute of Environ-
Immuno-spin trapping of DNA radicals: D. C. Ramirez, et al.; Nat. Methods 3, 123 mental Health Sciences, Research Triangle Park (RTP), NC, USA. firstname.lastname@example.org
(2006) Immuno-spin trapping of hemoglobin and myoglobin radicals derived from
nitrite-mediated oxidation: A. Keszler, et al.; Free Radic. Biol. Med. 40, 507 (2006)
Protein radical formation on thyroid peroxidase during turnover: M. Ehrenshaft,
et al.; Free Radic. Biol. Med. 41, 422 (2006) Protein radical formation on thyroid Benelux
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B1 radical formed by a dual role of xanthine oxidase and endothelial nitric oxide
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LIT: Immuno-spin trapping of DNA radicals: D. C. Ramirez, et al.; Nat.
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