β-Galactosidase Enzyme Assay System with Reporter Lysis Buffer

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed 3 × 10^5 cells	After 24 hours of transfection, perform assay and incubate for 30 min at 37 °C or until a faint yellow colour has developed.	Read the absorbance at 420nm

Upstream tips
Seed 3 × 10^5 cells
Protocol tips
After 24 hours of transfection, perform assay and incubate for 30 min at 37 °C or until a faint yellow colour has developed.
Downstream tips
Read the absorbance at 420nm

Publication protocol

In 6-well plates, 3 × 105 H1299 and Saos-2 cells were transfected in duplicate using Fugene transfection reagent with 0.5 μg/ml of pRGCΔFosLacZ plasmid (pRGC), containing the Ribosomal Gene Cluster (RGC), which includes three p53-binding sites with the same consensus sequence as the one of p53con, upstream β-galactosidase gene, and 2 μg/ml of pcDNA3-expression vector (empty, -TAp53, -Δ40p53). The following TAp53/Δp53 vector ratios were used: 0.5/0.5 and 0.5/1.5 µ/ml. The total amount of plasmid has been adjusted by adding pcDNA3-empty vector to reach 2.5 μg/ml of total transfected DNA in each condition. In addition, cell transfection efficiency was assessed by transfecting 1 μg/ml of a GFP-expressing vector. Twenty four hours post-transfection, cells were harvested and β-galactosidase activity was measured using β-galactosidase Enzyme Assay System kit (Promega).

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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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