Skip to Main content Skip to Navigation
New interface
Journal articles

A fast and reproducible cell- and 96-well plate-based method for the evaluation of P2X7 receptor activation using YO-PRO-1 fluorescent dye

Patrice Rat 1 Elodie OLIVIER 1 Caroline Tanter Anaïs Wakx Mélody Dutot 
1 UMR 8638 - Equipe C-TAC
COMETE - UMR 8638 - Chimie Organique, Médicinale et Extractive et Toxicologie Expérimentale
Abstract : The YO-PRO-1 assay provides a quantitative estimation of P2X7 receptor activation. P2X7 receptor is associated to pathological conditions including infectious, inflammatory, neurological, musculoskeletal disorders, pain and cancer. Most primary cells and cell lines from diverse origin may be used thanks to the ubiquitous distribution of P2X7 receptor. To study the activation of P2X7 receptor by chemicals or biological agents, we established a microplate-based cytometry protocol to accurately and rapidly quantify the activation of P2X7 receptor that leads to the formation of large pores in cell membranes. The YO-PRO-1 assay is based on the ability of cells to incorporate and bind YO-PRO-1 dye to DNA after activation of P2X7 receptor through pore formation. Cells are seeded in 96-well plates and incubated with the compound being tested for the appropriate time. The microplate is then incubated for 10 min with YO-PRO-1 staining solution. After the 10 min staining time, fluorescence signal is read using a microplate reader in 1 min. This procedure is easier and requires less handling steps than flow cytometry. 96-well plate based YO-PRO-1 assay is a reproducible and fast method to study both P2X7 receptor activation by toxic agents at subnecrotic concentrations and P2X7 receptor inhibition by antagonists.
Document type :
Journal articles
Complete list of metadata

https://hal-univ-paris.archives-ouvertes.fr/hal-03837761
Contributor : Elodie OLIVIER Connect in order to contact the contributor
Submitted on : Thursday, November 3, 2022 - 10:11:35 AM
Last modification on : Friday, November 4, 2022 - 3:58:05 AM

Links full text

Identifiers

Collections

Citation

Patrice Rat, Elodie OLIVIER, Caroline Tanter, Anaïs Wakx, Mélody Dutot. A fast and reproducible cell- and 96-well plate-based method for the evaluation of P2X7 receptor activation using YO-PRO-1 fluorescent dye. Journal of Biological Methods, 2017, 4 (1), pp.e64. ⟨10.14440/jbm.2017.136⟩. ⟨hal-03837761⟩

Share

Metrics

Record views

0