Streamline Your Research with the Transcreener ADP2 Assay
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2. Use of Enzyme at 40-85% of its saturating concentration (3 – 20% ATP conversion) produces good assay window Use of Enzyme at 60-85% of saturation allows accurate IC 50 determination directly from raw fluorescence data The assay accommodates [ATP] from 0.1 to 1,000 µM (ADP 2 Antibody concentration must be adjusted accordingly) The optimal [ADP 2 Antibody] can be calculated based on a linear equation: [Ab] = 1.08 [ATP] + 1 EC 60-85
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4. Determine IC 50 Values Directly from Raw Fluorescence Data via 0.4 Conversion Factor When enzyme reactions are set up appropriately, there is no need to run a standard curve We have shown empirically that IC 50 values determined from raw fluorescence data are directly proportional to those determined from product (ADP) formation
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![Fluorescence Polarization … Fluorescence Intensity … TR-FRET ,[object Object],[object Object],[object Object],[object Object]](https://image.slidesharecdn.com/transcreeneradpstreamlineyourresearch-110608140855-phpapp02/85/Streamline-Your-Research-with-the-Transcreener-ADP2-Assay-1-320.jpg)
![Use of Enzyme at 40-85% of its saturating concentration (3 – 20% ATP conversion) produces good assay window Use of Enzyme at 60-85% of saturation allows accurate IC 50 determination directly from raw fluorescence data The assay accommodates [ATP] from 0.1 to 1,000 µM (ADP 2 Antibody concentration must be adjusted accordingly) The optimal [ADP 2 Antibody] can be calculated based on a linear equation: [Ab] = 1.08 [ATP] + 1 EC 60-85](data:image/gif;base64,R0lGODlhAQABAIAAAAAAAP///yH5BAEAAAAALAAAAAABAAEAAAIBRAA7)