β-Galactosidase Enzyme Assay System with Reporter Lysis Buffer

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Add 150µl of Assay 2X Buffer to sample. Incubate the reactions at 37°C for 30 minutes or until a faint yellow color has developed.	Read the absorbance at 420nm.

Protocol tips
Add 150µl of Assay 2X Buffer to sample. 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

Transfection efficiency was assessed through cotransfection of a lacZ reporter construct, and β-galactosidase activity was determined using a commercially available β-galactosidase enzyme assay system according to the manufacturer's instructions (Promega, Madison, WI). Two to three independent transfections were performed for each experiment, and all samples were analyzed in triplicate for each transfection. For some experiments, transfection efficiency was further assessed on a single-cell level through cotransfection with a green fluorescent protein (GFP) construct and subsequent IFA. This procedure was performed using primary cells and HEK cells as a positive control, since the transfection efficiency of GFP for HEK cells approaches 90%. Using this method, we confirmed a HUVEC transfection efficiency of approximately 50% of cells in our experiments.

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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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