oxyluciferin+AMP+PPi+CO2+lightThis non-radioactive, homogeneous cell-based assay can be conveniently performed in microplates. The reagent is compatible with all culture media and liquid handling systems for high-throughput screening applications in 96-well and 384-well plates.Key Features:Safe. Non-radioactive assay (cf. 3H-thymidine incorporation assay).Sensitive and accurate. As low as 50 cells can be quantified.Homogeneous and convenient. Mix-incubate-measure type assay. No wash and reagent transfer steps are involved.Robust and amenable to HTS: Z’ factors of 0.6 to 0.7 are routinely observed in 96-well and 384-well plates. Can be readily automated on HTS liquid handling systems for processing thousands of samples per day.Applications:Cell proliferation: effects of cytokines, growth factor, nutrients.Cytotoxicity and apoptosis: evaluation of toxic compounds, anti-cancer antibodies, toxins, environmental pollutants etc.Drug discovery: high-throughput screening for anticancer drugs.Kit Contents:Assay Buffer, 1x10mlSubstrate, 1x120ulATP Enzyme, 1x120ulStorage and Stability:Store components at -20°C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Assay Procedure: In 96-Well Plates1. Cell Culture. Plate cells at 100ul/well in white opaque tissue culture plates. If desired, add 5ul test compounds and controls dissolved in PBS or culture medium per well. Rock plate lightly to mix and incubate for desired period of time (e.g. overnight).2. Assay. Bring all components to room temperature. Keep thawed ATP Enzyme on ice or 4°C. For each test well, mix 95ul Assay Buffer with 1ul Substrate and 1ul ATP Enzyme. Add 90ul Reconstituted Reagent to each test well and mix by tapping the plate. Incubate for 2 minutes at room temperature. Read luminescence on a luminometer. For most luminometers (Berthold Luminometer, LJL Analyst, Top Count, MicroBeta Counters, CLIPR and LeadSeeker), integration time of 0.1 to 5 sec is appropriate.Assay Procedure: In 384-Well Plates1. Cell Culture. Plate cells at 25ul/well in white opaque tissue culture plates. If desired, add 5ul test compounds and controls dissolved in PBS or culture medium per well. Rock plate lightly to mix and incubate for desired period of time (e.g. overnight).2. Assay. Bring all components to room temperature. Keep thawed ATP enzyme on ice or 4°C. For each test well, mix 30ul Assay Buffer with 0.3ul Substrate and 0.3ul ATP Enzyme. Add 25ul Reconstituted Reagent to each well and mix by tapping the plate. Incubate for 2 minutes at room temperature.Read luminescence on a luminometer. For most luminometers (Berthold Luminometer, LJL Analyst, Top Count, MicroBeta Counters, CLIPR and LeadSeeker), integration time of 0.1 to 5 sec is appropriate.General Considerations:Signal stability. After adding the Reconstituted Reagent, the luminescence signal is stable for about 15 min and decreases slow thereafter. Reading is best performed within 30 min.">
Adenosine 5'-triphosphate (ATP) is the chemical energy for cellular metabolism and is often referred to as “energy currency of the cell. ATP is produced only in living cells during photosynthesis and cellular respiration and consumed in cellular processes including biosynthetic reactions, motility and cell division. It is a key indicator of cellular activity and has been utilized as a measure of cell viability and cytotoxicity in research and drug discovery.Cytotoxicity Assay Kit provides a rapid method to measure intracellular ATP, cell viability and cytotoxicity. The single working reagent lyses cells to release ATP, which, in the presence of luciferase, immediately reacts with the Substrate D-luciferin to produce light. The light intensity is a direct measure of intracellular ATP concentration and hence number of living cells.ATP+D-luciferin+O2 ----luciferase--->oxyluciferin+AMP+PPi+CO2+lightThis non-radioactive, homogeneous cell-based assay can be conveniently performed in microplates. The reagent is compatible with all culture media and liquid handling systems for high-throughput screening applications in 96-well and 384-well plates.Key Features:Safe. Non-radioactive assay (cf. 3H-thymidine incorporation assay).Sensitive and accurate. As low as 50 cells can be quantified.Homogeneous and convenient. Mix-incubate-measure type assay. No wash and reagent transfer steps are involved.Robust and amenable to HTS: Z’ factors of 0.6 to 0.7 are routinely observed in 96-well and 384-well plates. Can be readily automated on HTS liquid handling systems for processing thousands of samples per day.Applications:Cell proliferation: effects of cytokines, growth factor, nutrients.Cytotoxicity and apoptosis: evaluation of toxic compounds, anti-cancer antibodies, toxins, environmental pollutants etc.Drug discovery: high-throughput screening for anticancer drugs.Kit Contents:Assay Buffer, 1x10mlSubstrate, 1x120ulATP Enzyme, 1x120ulStorage and Stability:Store components at -20°C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Assay Procedure: In 96-Well Plates1. Cell Culture. Plate cells at 100ul/well in white opaque tissue culture plates. If desired, add 5ul test compounds and controls dissolved in PBS or culture medium per well. Rock plate lightly to mix and incubate for desired period of time (e.g. overnight).2. Assay. Bring all components to room temperature. Keep thawed ATP Enzyme on ice or 4°C. For each test well, mix 95ul Assay Buffer with 1ul Substrate and 1ul ATP Enzyme. Add 90ul Reconstituted Reagent to each test well and mix by tapping the plate. Incubate for 2 minutes at room temperature. Read luminescence on a luminometer. For most luminometers (Berthold Luminometer, LJL Analyst, Top Count, MicroBeta Counters, CLIPR and LeadSeeker), integration time of 0.1 to 5 sec is appropriate.Assay Procedure: In 384-Well Plates1. Cell Culture. Plate cells at 25ul/well in white opaque tissue culture plates. If desired, add 5ul test compounds and controls dissolved in PBS or culture medium per well. Rock plate lightly to mix and incubate for desired period of time (e.g. overnight).2. Assay. Bring all components to room temperature. Keep thawed ATP enzyme on ice or 4°C. For each test well, mix 30ul Assay Buffer with 0.3ul Substrate and 0.3ul ATP Enzyme. Add 25ul Reconstituted Reagent to each well and mix by tapping the plate. Incubate for 2 minutes at room temperature.Read luminescence on a luminometer. For most luminometers (Berthold Luminometer, LJL Analyst, Top Count, MicroBeta Counters, CLIPR and LeadSeeker), integration time of 0.1 to 5 sec is appropriate.General Considerations:Signal stability. After adding the Reconstituted Reagent, the luminescence signal is stable for about 15 min and decreases slow thereafter. Reading is best performed within 30 min.