Functional genomic pore assay for mixed cell populations

摘要

A method of assaying nucleotide receptor P2X₇ pore activity in white blood cells contained within a mixed cell sample is provided comprising labeling white blood cells with a white blood cell-specific label; depolarizing the labeled white blood cells with an isotonic depolarizing solution; contacting the labeled white blood cells with dye and a P2X₇ agonist in an amount sufficient to activate P2X₇ pore activity; contacting the labeled white blood cells with a divalent cation in an amount sufficient to deactivate P2X₇ pore activity; and analyzing dye uptake whereby P2X₇ pore activity is quantified by the amount of dye taken up in labeled white blood cells treated with the P2X₇ agonist relative to labeled white blood cells in the absence of said P2X₇ agonist, said P2X₇ pore activity being corrected for sample age and by subtraction of P2X₇ pore activity contributed by nonviable white blood cells.