Cells were plated in 24 or 96 well plates and cultured for 24 hours. Dinaciclib, CFT-2718 or vehicle were diluted in cell culture medium and added to plates. For some assays, after 72 hours, CellTiter-Blue® (Promega, Fitchburg, WI) reagent was added to each well. After two hours incubation at 37°C, optical density readings were made in the 570 – 600 nm wave-length range, using a Perkin-Elmer ProXpress Visible-UV-fluorescence 16-bit scanner (Perkin-Elmer, Waltham, MA). In other experiments, a CellTiter-Glo® 2.0 luminescence Assay kit (Promega, Madison, WI) was used as an alternative means to assess viability, with data acquired on an EnVision™ Multilabel Reader (PerkinElmer, Santa Clara, CA, USA). In these assays, a time zero plate was incorporated into the assay to define GI50, TGI and LC50 (36 ).
Celltiter glo 2.0 luminescence assay kit
Manufactured by Promega
The CellTiter-Glo® 2.0 Luminescence Assay kit is a cell-based assay that quantifies the amount of ATP present in a sample, which is an indicator of metabolically active cells. The assay uses a thermostable luciferase to catalyze the reaction between ATP and luciferin, producing a luminescent signal that is proportional to the amount of ATP present.
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2 protocols using celltiter glo 2.0 luminescence assay kit
1
Cell Viability Assays for Drug Testing
2
Cell Viability Assay for Drug Evaluation
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Cells were plated in 24- or 96-well plates and cultured for 24 hours. Dinaciclib, CFT-2718, or vehicle were diluted in cell culture medium and added to plates. For some assays, after 72 hours, CellTiter-Blue (Promega) reagent was added to each well. After 2-hour incubation at 37°C, optical density readings were made in the 570- to 600-nm wavelength range, using a Perkin-Elmer ProXpress Visible-UV-fluorescence 16-bit scanner (Perkin-Elmer). In other experiments, a CellTiter-Glo 2.0 luminescence Assay Kit (Promega) was used as an alternative means to assess viability, with data acquired on an EnVision Multilabel Reader (PerkinElmer). In these assays, a time zero plate was incorporated into the assay to define GI50, TGI, and LC50 (36 ).
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