ONPG is a colorimetric and spectrophotometric substrate for detection of ß-galactosidase activity. ONPG is cleaved by ß-galactosidase via hydrolysis at the ß-1-4-glycosidic bond between 2-nitrophenol and galactose. The released 2-nitrophenol is measured spectrophotometrically at 405 nm. The absorbance intensity at this wavelength relates directly to the specific activity.
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