Rubidium efflux as a tool for the pharmacological characterisation of compounds with BK channel opening properties.
Methods Mol Biol
; 491: 267-77, 2008.
Article
en En
| MEDLINE
| ID: mdl-18998100
ABSTRACT
This chapter describes a method of assaying rubidium (Rb(+)) efflux as a measure of potassium channel activity. In this assay, rubidium acts as a tracer for potassium movement across the cell membrane. HEK 293 cells expressing the alpha subunit of the human brain large-conductance, voltage-activated, calcium-sensitive potassium channel (BK channel) are loaded with Rb(+), washed, and then incubated under experimental conditions. The cell supernatant is removed, and the remaining cell monolayer lysed. These two samples contain Rb(+) that has moved out of the cell and Rb(+) that remains in the cell, respectively. Measurement of the Rb(+) content of these samples by flame atomic absorption spectrometry allows calculation of the percentage Rb(+) efflux and, depending on the experimental design, provides pharmacological data about the control and test compounds used. In this chapter, we describe the protocol and steps for optimisation and illustrate this with data obtained using NS1619, a well-characterised BK channel opener.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Canales de Potasio de Gran Conductancia Activados por el Calcio
Límite:
Humans
Idioma:
En
Revista:
Methods Mol Biol
Asunto de la revista:
BIOLOGIA MOLECULAR
Año:
2008
Tipo del documento:
Article
País de afiliación:
Reino Unido