Wisconsin Alumni Research Foundation

Kathleen Schell

Paul J Bertics

Kirk Jeffrey Hogan

Loren C Denlinger

A method of assaying nucleotide receptor P2Xpore 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 P2Xagonist in an amount sufficient to activate P2Xpore activity; contacting the labeled white blood cells with a divalent cation in an amount sufficient to deactivate P2Xpore activity; and analyzing dye uptake whereby P2Xpore activity is quantified by the amount of dye taken up in labeled white blood cells treated with the P2Xagonist relative to labeled white blood cells in the absence of said P2Xagonist, said P2Xpore activity being corrected for sample age and by subtraction of P2Xpore activity contributed by nonviable white blood cells.

Functional genomic pore assay for mixed cell populations

A method of rapidly assaying nucleotide receptor P2Xpore activity in white blood cells contained within a blood sample. A method according to the invention includes the steps of: (a) labeling the white blood cells with a white blood cell-specific label; (b) depolarizing the labeled white blood cells with an isotonic depolarizing solution; (c) contacting the labeled white blood cells with a dye and a P2Xagonist in an amount sufficient to activate nucleotide receptor P2Xpore activity; (d) contacting the labeled white blood cells with a divalent cation in an amount sufficient to deactivate nucleotide receptor P2Xpore activity; and (e) analyzing dye uptake in labeled white blood cells whereby nucleotide receptor P2Xpore activity is quantified by the amount of dye taken up in labeled white blood cells treated with P2Xagonist relative to labeled white blood cells in the absence of P2Xagonist.

Growing community of inventors

Madison, WI, United States of America

Kathleen Schell