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ATP Assay, Bioluminescence is a one-step, 20-minute assay that measures ATP to monitor the number of viable mammalian cells in cell proliferation, cytotoxicity, and other cell-based assays. This assay uses firefly luciferase, a 61 kDa monomeric enzyme, responsible for catalyzing the oxidation of firefly luciferin. This reaction produces a stable and long-lasting (> 2 hours) light emission of 560 nm, which directly corresponds to the concentration of ATP present in the cells. This assay can reliably detect as few as 50 cells per well and is free of unpleasant odors, as the firefly luciferase does not require enzymatic stabilization with dithiothreitol (DTT). ATP Assay, Bioluminescence is supplied as a single, ready-to-use reagent (no washing of cells required) and can be utilized in 96- or 384-well microplates, making it an ideal solution for high-throughput screening experiments. Quantification can be carried out using a luminescent capable microplate reader (luminometer).
Figure 1. ATP Assay, Bioluminescence Shows Luminescent Signal Output Is Directly Proportional to Cell Number
(A) HEK293 cells were plated in a white opaque 96-well plate with two-fold serial dilution at a density of 50 - 50 x 10^3 cells per well. Luminescent output was measured 20 minutes after addition of ATP Assay, Bioluminescence and shows a linear relationship (r2 = 0.99) between cell number and luminescence signal over three orders of magnitude and can detect as low as 50 cells per well. Values are representative of three replicates per cell number. (B) ATP Assay, Bioluminescence shows a linear relationship (r2 = 0.99) between cell number per well and luminescence signal even at low seeding densities (100 - 1600 cells).
Figure 2. ATP Assay, Bioluminescence has a Stable Luminescent Signal with a Half-Life of More than 2 Hours
HEK293 cells were plated in a white opaque 96-well plate at a density of 25 x 10^3 cells per well. After addition of ATP Assay, Bioluminescence, plate was incubated at room temperature. Luminescent output was measured 20 minutes after reagent addition and additional luminescence measurements were recorded at 60, 120, 180, 240, and 300 minutes. The graph shows ATP Assay, Bioluminescence half-life of the luminescent signal is more than 2 hours.
