Oxidative stress resulting in the formation of biological radicals has been implicated in a number of human diseases, such as cancer and aging. There is, however, a paucity of reliable methods for in vivo or ex vivo detection of radical formation. Until now the only general technique that allowed for the detection of these highly reactive species was electron spin resonance (ESR) using spin traps. One of the most popular of these spin traps is 5,5-dimethyl-1-pyrroline N-oxide (DMPO). In the ESR method, radicals are trapped by DMPO, and the DMPO spin adduct signal is measured quantitatively by an ESR spectrometer. The DMPO, mAb (1) (Prod. No. ALX-803-340) specifically reacts with DMPO-protein and DMPO-DNA adducts and therefore highly simplifies the detection of radical formation compared to the ESR method.
Product Details
| Alternative Name: | 5,5-dimethyl-1-pyrroline-N-oxide |
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| Clone: | 1 |
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| Host: | Mouse |
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| Isotype: | IgG |
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| Immunogen: | DMPO-octanoic acid conjugated to ovalbumin. |
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| Source: | Hybridoma tissue culture supernatant. |
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| Species reactivity: | Species independent
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| Specificity: | Recognizes DMPO (Prod. No. ALX-430-090), DMPO-octanoic acid, DMPO-protein adducts and DMPO-DNA adducts. |
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| Crossreactivity: | Does not cross-react with non-adducted proteins or DNA. Does not cross-react with ovalbumin. |
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| Applications: | ELISA, IP, WB
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| Recommended Dilutions/Conditions: | Western Blot (1-10ug/ml)
ELISA (1-10ug/ml)
Suggested dilutions/conditions may not be available for all applications.
Optimal conditions must be determined individually for each application. |
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| Purity Detail: | Protein G-affinity purified. |
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| Formulation: | Liquid. In PBS, pH 7.4, containing 10% glycerol and 0.02% sodium azide. |
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| Handling: | Dilute only prior to immediate use. |
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| Shipping: | Blue Ice |
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| Long Term Storage: | +4°C |
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| Regulatory Status: | RUO - Research Use Only |
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Figure: Western Blot analysis using DMPO, mAb (1) (Prod. No. ALX-803-340).Method:Samples are 2.5 µg each from reactions containing:1= 10 µM Mb only2= 10 µM Mb + 100 mM DMPO3= 10 µM Mb + 100 µM H2O2 + 100 mM DMPO4= 10 µM Mb + 10 µM H2O2 + 10 mM DMPOWestern conditions are:-either 0.1 or 1.0 µg/ml anti-DMPO IgG for 1 hr at rt in 1XPBS/0.5% Tween-wash 4X in 1XPBS/0.5% Tween-1:10,000 anti-mouse IR800 for 1 hr at rt in 1XPBS/0.5% Tween-wash 2X in 1XPBS/0.5% Tween, wash 2X in PBS-rinse in H2O-Scan on Licor Odyssey
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Product Literature References
Melanosomes in pigmented epithelia maintain eye lens transparency during zebrafish embryonic development: M. Takamiya, et al.; Sci. Rep.
6, 25046 (2016),
Application(s): Immuno spin trapping assay with DMPO,
Abstract;
Full Text
Immuno-spin trapping of a post-translational carboxypeptidase B1 radical formed by a dual role of xanthine oxidase and endothelial nitric oxide synthase in acute septic mice: S. Chatterjee, et al.; Free Radic. Biol. Med.
46, 454 (2009),
Abstract;
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