Assay Method Information

Assay Name:  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 L 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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