|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!