|Method:||Colorimetric assay, 12x8-well strips|
|Sample type:||Serum, EDTA plasma, biological fluids|
|Standard range:||1-point calibration, 2 controls|
|Sample size:||10 µl / well|
|Incubation time:||15 min|
LOD: 7 µmol/l (0 µg/ml + 3 SD)
Intra-assay (n=12) ≤ 3%, Inter-assay (n=12) ≤ 5%
Values from apparently healthy individuals:
Median = 372 µmol /l (serum < 350 µmol/l, EDTA-plasma <400 µmol/l)
It is recommended to establish the normal range for each laboratory.
Principle of the assay:
The peroxide concentration is determined by reaction of the biological peroxides with peroxidase and a subsequent colour-reaction using TMB as substrate. After addition of a stop solution, the coloured liquid is measured photometrically at 450 nm.
A calibrator is used to calculate the concentration of circulating biological peroxides in the sample (one-point calibration).
Manual ELISAs - easily adaptable for automation!
INSTRUCTIONS FOR USE
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Cells and tissues are sensitive to oxidative stress, caused by the formation of free radicals. If not deactivated by antioxidants, organic peroxides and hydroperoxides are the first reaction products between cellular constituents and free radicals or other reactive oxygen derivates.
The determination of the oxidative status / oxidative stress is essential in today's medical research and diagnostics. Methods used so far were either expensive (HPLC), or detected only degradation products of polyunsaturated fatty acids, like TBARS (thiobarbituric acid reactive substances).
The Biomedica OxyStat assay measures the total peroxide concentration of a sample, utilizing a quick and simple assay procedure. Results show a direct correlation between free radicals and circulating biological peroxides and thus allow the characterization of the oxidative status in biological samples.
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