title: A homogeneous assay to assess GABA transporter activity.
abstract: This unit describes a convenient functional uptake assay for GABA transport into cell lines transiently transfected with GABA transporter-1 (GAT-1) and other GAT isoforms. This facile, homogeneous assay allows for the determination of K(m), V(max), and K(i) values. The assay utilizes commercially available microtiter plates that contain scintillant embedded in the bottom of the wells. Whereas a signal is generated as the cell accumulates the labeled neurotransmitter, label in the medium is undetected. While GABA uptake is observed in several cell lines transfected with GAT-1, K(m) values for GABA uptake may vary with the cell line. This indicates that the choice of cell line is an important consideration when conducting uptake assays.
authors.affiliation: Cephalon Inc., West Chester, Pennsylvania, USA.
authors.forename: Karla K
authors.initials: KK
authors.lastname: Kopec
authors.forename: Beth Ann
authors.initials: BA
authors.lastname: McKenna
authors.forename: Daniel
authors.initials: D
authors.lastname: Pauletti
mesh_terms: D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D050485:GABA Plasma Membrane Transport Proteins / Q000032:analysis* / Q000502:physiology; D057809:HEK293 Cells; D006801:Humans; D005680:gamma-Aminobutyric Acid / Q000378:metabolism*
chemical_list: D050485:GABA Plasma Membrane Transport Proteins; D005680:gamma-Aminobutyric Acid