Description
The CellTiter-Glo® One Solution Assay is a homogeneous method of determining the number of viable cells in culture based on quantitation of the ATP present, which indicates the presence of metabolically active cells. This frozen, ready-to-use format is based on the original CellTiter-Glo® Luminescence Cell Viability Assay chemistry and eliminates the need to combine buffer with lyophilized substrate when preparing reagent. The CellTiter-Glo® Assay is designed for use with multiwell-plate formats, making it ideal for automated high-throughput screening (HTS) in 96- to 1536-well format, and cell proliferation and cytotoxicity assays.
The homogeneous "add-mix-measure" format results in cell lysis and generation of a luminescent signal proportional to the amount of ATP present. The amount of ATP is directly proportional to the number of cells present in culture. The CellTiter-Glo® One Solution Assay generates a stable "glow-type" luminescent signal with a half-life of greater than three hours. This extended half-life eliminates the need for reagent injectors and provides flexibility for continuous or batch-mode processing of multiple plates.
No reagent preparation is required; simply thaw and "add-mix-measure". This format reduces the number of plate-handling steps. Data can be recorded 10 minutes after reagent addition, and the assay measures cells at numbers below the detection limits of standard colorimetric and fluorometric assays.
Volumes are convenient for HTS applications, and the assay can be used with various multiwell formats including 96-well, regular, or low volume 384- and 1536-well plates. Data can be recorded by luminometer or CCD camera imaging device.
The CellTiter-Glo assay provides a stable, luminescent signal with a half-life >3 hours, depending on cell type and culture medium used, and it can be used with other nonlytic compatible assay chemistries from Promega.
Notes
Cat.# G8461 is sufficient for 1,000 assays at 100µl/assay in 96-well plates or 4,000 assays at 25µl/assay in 384-well plates. Cat.# G8462 is sufficient for 5,000 assays at 100µl/assay in 96-well plates or 20,000 assays at 25µl/assay in 384-well plates.
Cell number correlates with luminescent output.
A direct relationship exists between luminescence measured with the CellTiter-Glo® One Solution Assay and the number of cells in culture over three orders of magnitude. Serial twofold dilutions of Jurkat cells were made in RPMI 1640 with 10% FBS and plated in three formats using automated dispensing equipment. Volumes plated were 100µl of cells per well in a 96-well plate ( Panel A ), 10µl of cells per well in low-volume 384-well plate ( Panel B ) and 4µl of cells per well in a 1536-well plate ( Panel C ). An equivalent volume of CellTiter-Glo® One Solution was dispensed into each well. Luminescence was recorded 10 minutes after addition of CellTiter-Glo® One Solution. There is a linear relationship (r 2 > 0.99) between luminescent signal and cell number in each plate format.
