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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 is a convenient method
for assaying β-galactosidase activity in lysates prepared from cells transfected
with β-galactosidase reporter vectors such as the 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 β-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.
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