Standardized method to minimize variability in a functional P2X(7) flow cytometric assay for a multi-center clinical trial.

ABSTRACT:

Background

Flow cytometric analysis of human P2X(7) pore activity segregates variant from common P2RX7 genotypes and may serve as a biomarker for cancer, pain, inflammation, and immune responses to infection. Standardization is needed to accommodate variable sample age and instrumentation differences in a multicenter clinical trial.

Methods

CD14-PE-stained whole blood samples were treated with YO-PRO-1 combined with a P2X(7) agonist (BzATP) or control, followed by the addition of PI after closure of the P2X(7) pore. Recalled instrument settings from previous publications were used to adapt a standardized fluorescent particle-adjusted set-up method. Experiments were performed to compare the two methods while evaluating components of systematic variability and facilitating rel

SUBMITTER: Korpi-Steiner NL 

PROVIDER: S-EPMC2622726 | biostudies-literature | 2008 Sep

REPOSITORIES: biostudies-literature