El diclorhidrato de o-fenilendiamina (OPD) es un sustrato cromógeno que puede utilizarse en procedimientos de enzimoinmunoanálisis de absorción (ELISA), ya que utiliza conjugados de peroxidasa de rábano picante. El sustrato rinde un producto final soluble que es de color naranja-marrón y puede leerse en el espectrofotómetro a 450 nm. La reacción del OPD puede detenerse con HCl 3N o H2SO4 3 M y leerse a 492 nm.
o-Phenylenediamine (o-PD) is a colorless and nonfluorescent molecule.[1]
Aplicación
o-Phenylenediamine dihydrochloride has been used as a substrate for horseradish-peroxidase-conjugated secondary antibody in enzyme-linked immunosorbent assay (ELISA).[2][3][4]
Reconstitución
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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NBT-BCIP substrate system aids in western blotting and immunohistological staining, producing a blue-purple insoluble end product.
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