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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.
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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
| Regulatory Status |
RUO – Research Use Only |
|---|
Related Products
| Alternative Name | 5,5-Dimethyl-1-pyrroline-N-oxide, 2,2-Dimethyl-3,4-dihydro-2H-pyrrole 1-oxide |
|---|---|
| CAS | 3317-61-1 |
| Purity | >98% (HPLC) |