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Adenosine Diphosphate/Adenosine Triphosphate Ratio (ADP/ATP), Assay Kit, BioAssay™
Adenosine Diphosphate/Adenosine Triphosphate Ratio (ADP/ATP), Assay Kit, BioAssay™
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| Catalog # | A0014-98 | | Description | Changes in the ADP/ATP ratio have been used to differentiate modes of cell death and viability. Increased levels of ATP and decreased levels of ADP signify proliferating cells. Conversely, decreased levels of ATP and increased levels of ADP represent apoptotic or necrotic cells where the decrease in ATP and increase in ADP is much more pronounced in necrosis versus apoptosis. ADP/ATP Ratio Assay Kit provides a rapid method to measure ADP and ATP levels for the screening of apoptosis, necrosis and cell proliferation in mammalian cells. The assay involves two steps. In the first step, the working reagent lyses cells to release ATP and ADP. In the presence of luciferase, ATP immediately reacts with the Substrate Dluciferin to produce light. The light intensity is a direct measure of intracellular ATP concentration. Luciferase ATP + D-luciferin + O2 oxyluciferin + AMP + PPi + CO2 + light In the second step, the ADP is converted to ATP through an enzyme reaction. This newly formed ATP then reacts with the D-luciferin as in the first step. Due to a special formulation of the reagent system which greatly stabilizes the light signal generated by the luciferase reaction, the luminescence from the initial ATP measurement remains stable throughout this assay. Therefore, the second light intensity measured represents the total ADP and ATP concentration in the sample. This non-radioactive, homogeneous cell-based assay is performed in microplates. The reagent is compatible with all culture media and with all liquid handling systems for high-throughput screening applications in 96- well and 384-well plates. | | Key Features | Safe. Non-radioactive assay. Homogeneous and convenient. "Mix-incubate-measure" type assay. No wash and reagent transfer steps are involved. Robust and amenable to HTS: Z' factors of 0.5 and above 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 | Apoptosis and Necrosis determination in cells. Cell proliferation: effects of cytokines, growth factor, nutrients. Drug discovery: high-throughput screening for anticancer drugs. | | Kit Contents | Assay Buffer: 10ml Substrate: 120ml Cosubstrate 120ml ATP Enzyme: 120ml ADP Enzyme: 120ml Storage conditions: store all reagents at -20°C. This product is shipped on dry ice. Shelf life of at least 6 months. Precautions: reagents are for research use only. Normal precautions for laboratory reagents should be exercised while using the reagents. Please refer to Material Safety Data Sheet for detailed information. | | Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
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