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Abstract for β-Galactosidase Enzyme Assay System with Reporter Lysis Buffer
β-Galactosidase is a commonly used reporter molecule. The β-Galactosidase Enzyme Assay System with Reporter Lysis Buffer (Cat.# E2000) is a convenient method for assaying β-galactosidase activity in lysates prepared from cells transfected with β-galactosidase reporter vectors such as Promega's pSV-β-Galactosidase Control Vector.
The standard assay is performed by adding a dilute sample to an equal volume of Assay 2X Buffer, which contains the substrate ONPG (o-nitrophenyl-beta-D-galactopyranoside). Samples are incubated for 30 minutes, during which time the Beta-Galactosidase hydrolyzes the colorless substrate to o-nitrophenyl, which is yellow. The reaction is terminated by addition of sodium carbonate, and the absorbance is read at 420nm with a spectrophotometer.