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The Beta-Galactosidase Enzyme Assay System with Reporter Lysis Buffer is a convenient method for assaying beta-galactosidase activity in lysates prepared from cells transfected with beta-galactosidase reporter vectors such as the pSV-Beta-Galactosidase Control Vector. The standard assay is performed by adding a dilute sample to an equal volume of Assay 2X Buffer that contains the substrate ONPG (o-nitrophenyl-beta-D-galactopyranoside). Samples are incubated for at least 30 minutes, during which time the Beta-Galactosidase hydrolyzes the colorless substrate to o-nitrophenyl, which is yellow. The reaction can be terminated by addition of sodium carbonate, and the absorbance at 420mM is measured by spectrophotometry.
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