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78751644429
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2+ can flow through the open channel and stimulate intracellular fluorescence. Conversely, if the TRPV1 channel is exposed to the agonist capsaicin, in the presence of antagonism from the test compound, the channel will remain closed and intracellular fluorescence will not occur. In this way the same TRPV1 transfected cell line and assay can be used to search for both agonists and antagonists. Results are an average of at least two independent experiments with three replicates at each concentration.
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48849099961
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W.S. Cheung, R.R. Calvo, B.A. Tounge, S.-P. Zhang, D.R. Stone, M.R. Brandt, T. Hutchinson, C.M. Flores, and M.R. Player Bioorg. Med. Chem. Lett. 18 2008 4569
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78751648075
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Aqueous solubilities were determined using a medium-throughput adaptation of a shake-flask methodology. A 10 mM solution of the test compound in DMSO was added to 0.05 M phosphate buffered saline pH 7.4 such that the final concentration of DMSO was 2%. The resultant mixture was then vortex mixed (1500 rpm) for 24 ± 0.5 h at 21 ± 2 °C. After mixing, the resultant solution/suspension was filtered under vacuum using a filter plate (Millipore Multiscreen HTS, 0.4 μM). The concentration of the compound in the filtrate was determined by High Performance Liquid Chromatography (HPLC) running a generic acid gradient method with UV detection at 230 nm. Peak areas from analysis of the diluted filtrates were quantified by comparison to a calibration line prepared by injecting onto the HPLC three different volumes of a 50 μM solution of the test compound in DMSO. Solubilities were determined in duplicate for each test compound and average values reported.
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78751646352
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PERCH software Version 1/2005, PERCH Solutions Ltd, Kuopio, Finland
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PERCH software Version 1/2005, PERCH Solutions Ltd, Kuopio, Finland.
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0035425705
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note
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Methodology adapted from Vellani, V.; Mapplebeck, S.; Moriondo, A.; Davis, J. B.; McNaughton, P. A. J. Physiol. 2001, I, 813-825. Primary cultures of rat dorsal root ganglion neurones were prepared from adult Wistar rats. Standard whole-cell patch clamp recordings were made from neurones on days 2-4 in culture. The neurones were exposed to capsaicin (500 nM) applied through a local perfusion system for 1 s, repeated every 30 s. Capsaicin evoked an inward current which typically showed a brief period of runup in amplitude before stabilizing. Once the amplitude was stable, test compounds were applied through the local perfusion system. Compounds were applied until the amplitude was judged to have plateaued over at least three successive capsaicin applications.
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