ATPlite一步法检测试剂盒.pdf

Introduction A number of strategies are currently being implemented in an effort to increase the success rate in identifying potential therapeutic lead compounds and reducing the number of late stage clinical trial failures. One approach is to perform ADME/Tox profiling at an earlier stage in the drug discovery process. The ATPlite? 1step Luminescence Assay System has been used to perform automated cytotoxicity assays on the JANUS? Cellular Workstation creating an easy-to-use, accurate integrated walk-away system for cellular assays. ATPlite 1step assay is a homogeneous, luciferase-based luminescence detection assay system for the quantitative measurement of proliferation and cytotoxicity of cultured mammalian cells. ATP level is a general marker for determin- ing cell viability, as it is present in all metabolically active cells and its concentration declines rapidly when cells undergo necrosis or apoptosis. Model cytotoxicity assays have been performed using both U937 suspension cells, and HepG2 adherent cells. Resulting cell viability was measured following treatment with either Staurosporine or Actinomycin D. Cell viability was determined as a function of both the time of exposure and the concentration of the test compounds. Utilization of the ATPlite 1step Detection System for Homogeneous Automated Cytotoxicity Assays on the JANUS Cellular Workstation A P P L I C A T I O N N O T E Authors Steve Hurt Hanh Le PerkinElmer, Inc. 940 Winter Street Waltham, MA 02451 USA Figure 1. The JANUS Cellular Workstation is a fully integrated system solution providing liquid handling, detection, plate movement, capacity as well as scheduling, incubation and plate washing options. Figure 2. Typical JANUS deck layout for the ATPlite 1step assay. The reagent reservoir is highlighted here in green. Materials and methods The Cellular Workstation PerkinElmer’s JANUS Cellular Workstation was utilized for the ATPlite 1step assay. The Workstation was equipped with a 384-channel Mod