β-Galactosidase Enzyme Assay System with Reporter Lysis Buffer

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed 1 × 10^5 cells per well	Add 150µl of Assay 2X Buffer to samples. Incubate the reactions at 37°C for 30 minutes or until a faint yellow color has developed	Read the absorbance at 420nm

Upstream tips
Seed 1 × 10^5 cells per well
Protocol tips
Add 150µl of Assay 2X Buffer to samples. Incubate the reactions at 37°C for 30 minutes or until a faint yellow color has developed
Downstream tips
Read the absorbance at 420nm

Publication protocol

CHO-K1 cells were seeded in a 24-well plate (1 × 105 cells per well) and incubated overnight before being transfected using Metafectene (Biontex, Munich, Germany) with 0.3 μg each of PPARα expression vector and DR-1 (PPAR recognition site) luciferase reporter plasmids and 20 ng of a β-galactosidase reporter plasmid. Transfected cells were incubated for 24 h and then treated with linalool or fenofibrate for 24 h. The cells were lysed, and luciferase activity was measured with the Firefly Luciferase Assay Kit (Biotium, Hayward, CA) using a luminometer (Victor3, PerkinElmer, Waltham, MA). Relative promoter activity was computed by normalization to β-galactosidase activity, as determined with the β-Galac­tosidase Enzyme Assay System (Promega Corp., Madison, WI).

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Manufacturer protocol

Download the product protocol from Promega for β-Galactosidase Enzyme Assay System with Reporter Lysis Buffer below.

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