Enabling a high efficiency assay for GABA isoforms using transient transfection

Presented:
2011-12-15

Speaker:
Leggy Arnold PhD,
University of Wisconsin

Abstract

IonFlux is the only high throughput automated patch clamp system with continuous perfusion and no delay between compound additions.
Learn how the Arnold lab at University of Wisconsin, Milwaukee has developed a successful assay using the IonFlux 16 for determining GABA-A pharmacology.

What will be covered:

  • Overview of the IonFlux system
  • Assay development process for using transient transfection with the IonFlux system
  • Key factors in optimizing assay performance and key performance metrics
  • Validation data and developed capability for determining subunit-dependent pharmacology


More about the Webinar and Prof. Arnold's Research...
GABA-A receptors are ligand-gated ion channels found in the synapses of neurons that conduct chloride ions across neuronal cell membranes. The receptor contains two binding sites for γ-aminobutyric acid (GABA), the primary inhibitory neurotransmitter in the central nervous system. There are at least nineteen different individual GABA-A receptor subunits that assemble the pentameric structure in different individual combinations to form the native receptor (α1-6, β1-3, γ1-3, δ, ρ1-3, and minor subunits). We determined the changes in Cl- flux in the presence of GABA for HEK293T cells transfected with different α subunits and β3γ2 subunits using the IonFlux system. Additionally, we determined the modulation of Cl- flux by benzodiazepine Hz166. The comparison with manual electrophysiological measurements using the same subtype plasmids showed that meaningful pharmacological measurements are acquired with the IonFlux instrument. We will also present technical data including timing, success rate, cell preparation and inter-experiment repeatability.