Not available outside of the UK & Ireland.
Application
o-Phenylenediamine Free Base is a peroxidase substrate suitable for use in ELISA procedures. The substrate produces a soluble end product that is orange-brown in color and can be read spectrophotometrically at 450 nm. The OPD reaction may be stopped with 3 N HCl or 3 M H2SO4 and read at 492 nm.
o-Phenylenediamine has been used to detect transforming growth factor (TGF-β) and to measure anti-cTfRMAb-ScFv fusion protein using enzyme linked immunosorbent assay (ELISA).
Biochem/physiol Actions
o-Phenylenediamine (o-PD) is a potent substrate for spectrophotometric horseradish peroxidase (HRP)-mediated enzyme-linked immunosorbent assay (ELISA), which gives 2,3-diaminophenazine (DAP) as a product of o-PD oxidation.
Reconstitution
Dissolve one tablet in 0.05 M phosphate-citrate buffer, pH 5.0, to the desired concentration (typically an OPD concentration of 0.4 mg/ml is used). Add 40 µl of fresh 30% hydrogen peroxide per 100 ml of substrate buffer solution, immediately prior to use. Tablets are individually packaged in foil packets.
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