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| | Ethidium Bromide Uptake Assay |
| Description: | hP2X7-expressing HEK 293 cells were re-suspended at 2.5 × 10^6 cells/mL in assay buffer composed of 10 mM HEPES, 5 mM N-methyl-D-glutamine, 5.6 mM KCl, 10 mM D-glucose, and 0.5 mM CaCl2 (pH 7.4) supplemented with either 280 mM sucrose or 140 mM NaCl, and an 80 µL aliquot was added to each well of a 96-well plate. To each compound were added and KN-62 as the positive control, followed by the hP2X7R agonist benzoyl ATP (BzATP). The plates were incubated at 37 °C for 2 h and the cellular accumulation of ethidium ion was determined by measuring fluorescence (excitation = 530 nm; emission = 590 nm). |
| Affinity data for this assay | |
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