β-Galactosidase Enzyme Assay System with Reporter Lysis Buffer

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 that contains the substrate ONPG (o-nitrophenyl-β-d-galactopyranoside). Samples are incubated for at least 30 minutes, during which time the β-Galactosidase hydrolyzes the colorless substrate to o-nitrophenyl, which is yellow. The reaction may be terminated by addi...

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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 that contains the substrate ONPG (o-nitrophenyl-β-d-galactopyranoside). Samples are incubated for at least 30 minutes, during which time the β-Galactosidase hydrolyzes the colorless substrate to o-nitrophenyl, which is yellow. The reaction may be terminated by addition of sodium carbonate, and the absorbance at 420nm is measured by spectrophotometry.

Features - Benefits

• Safe: Non-isotopic assay.
• Versatile: The assay can be used in a 96-well plate format.
• Flexible: Reporter Lysis Buffer allows firefly luciferase, CAT and β-galactosidase assays to be performed from the same cell extract.

Applications

• This system is appropriate for measuring β-galactosidase enzyme activity in mammalian or bacterial cell extracts.

Notes

Cat.# E2000 contains sufficient reagents for 65 standard assays or 200 assays in a 96-well plate format.

References

1. Miller, J.H., ed. (1972) Experiments in Molecular Genetics, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
2. Sambrook, J. et al. (1989) Molecular Cloning, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
3. Schenborn, E. and Goiffon, V. (1993) Promega Notes 41, 11–3.

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