β-Gal Assay Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Assay cells after 24h transfection. Add 70 µl of ONPG and 200 µl 1X Cleavage Buffer with β-mercaptoethanol. Incubate samples at 37°C for 30 minutes	Read the absorbance at 420 nm

Protocol tips
Assay cells after 24h transfection. Add 70 µl of ONPG and 200 µl 1X Cleavage Buffer with β-mercaptoethanol. Incubate samples at 37°C for 30 minutes
Downstream tips
Read the absorbance at 420 nm

Publication protocol

For transfection, Panc-1, Aspc-1, and Bxpc-3 cells were seeded at 5 × 104 cells per well in 24-well plates. The β-galactosidase plasmid (50 ng) and pGL3-FBP1- Luc reporter (200 ng), along with a pcDNA3.1-Flag-NPM1 plasmid, were transiently transfected into the cells. Twenty-four hours post-transfection, cells were harvested and lysed. The luciferase and β-galactosidase activities were measured with a luciferase reporter gene assay kit (Invitrogen) and a β-galactosidase kit (Invitrogen), respectively. The transfection efficiency across plates was normalized to β-galactosidase activity, and all transfections were repeated three times in duplicate.

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Manufacturer protocol

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