Il cloridrato di o-fenilendiammina (OPD) è un substrato cromogeno adatto all'uso nel dosaggio con immunoadsorbente legato a un enzima (ELISA) in quanto si avvale di coniugati a perossidasi di rafano Il substrato sviluppa un prodotto finale solubile arancio-bruno e può essere letto con spettrofotometria a 450 nm. La reazione dell′OPD può essere arrestata con HCl 3N o H2SO4 3 M e può essere letta a 492 nm.
o-Phenylenediamine dihydrochloride is a binucleophile and its condensation with α,β-unsaturated carbonyl compounds and aldehydes yields benzimidazoles.[1][2]
Applicazioni
o-Phenylenediamine dihydrochloride has been used as a substrate for horse radish peroxidase in detecting:
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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The reaction of o-phenylenediamine with alpha, beta-unsaturated carbonyl compounds
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NBT-BCIP substrate system aids in western blotting and immunohistological staining, producing a blue-purple insoluble end product.
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