RealTime-Glo™ MT Cell Viability Assay

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Add RealTime-Glo reagents and incubate at 5% CO2 and 37°C every 30 min for 24 h	Luminescence was determined every 15 min for 5 h

Protocol tips
Add RealTime-Glo reagents and incubate at 5% CO2 and 37°C every 30 min for 24 h
Downstream tips
Luminescence was determined every 15 min for 5 h

Publication protocol

A549 cells (1,000/well) were plated in a 384-well plate in 40 μL of media. The cells were incubated overnight in a 37°C, 5% CO2-humidified atmosphere. The media was removed and replaced with 72 μL of CO2-independent media (Gibco) containing real-time cell viability reagents (RealTime-Glo MT Cell Viability Assay). The plate was read on a Tecan M200 plate reader set at 37°C every 30 min for 24 h. At 24 h, 8 μL of 2,000-μg/mL digitonin in CO2-independent media was added to the cells, and the luminescence was determined every 15 min for 5 h. The A549 cell line was chosen to represent a commonly used adherent cancer cell line.

Fresh rat hepatocytes were obtained from Bioreclamation IVT and plated in 72-μL InVitroGRO CP Medium containing the RealTime-Glo reagents. The plate was read on a Tecan M200 plate reader with gas control module (GCM) set at 5% CO2 and 37°C every 30 min for 24 h. At 24 h, 8 μL of 2,000-μg/mL digitonin in InVitroGRO CP Medium was added to the cells, and the luminescence was determined every 15 min for 5 h. Hepatocytes were chosen to represent a commonly used nonproliferating primary cell line.

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Manufacturer protocol

Download the product protocol from Promega for RealTime-Glo™ MT Cell Viability Assay below.

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