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| | Ca2+ Influx Functional Assay for the Determination of in Vitro Activity. |
| Description: | The functional antagonist activity of compounds at the TRPV1 receptor was determined with a Ca2+ influx assay by measuring the effect on capsaicin (0.05 uM) evoked increase in intracellular Ca2+ levels using cells stably expressing recombinant human TRPV1. All compounds were tested over an 11-point half-log concentration range. The fluorescent Ca2+ chelating dye fluo-4 was used as an indicator of the relative levels of Ca2+ in a 96-well format using a fluorescence imaging plate reader. Fluorescence readings were made at 1-5 s intervals over the 240 s course of the experimental run. The peak increase in relative fluorescence units was calculated and expressed as a percentage of the 0.05 uM capsaicin (control) response. Curve fits of the data were solved using a four-parameter logistic Hill equation in GraphPad Prism, and IC50 values were calculated. |
| Affinity data for this assay | |
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