UCPH-101

Cell experiment:

Cells are split into poly-D-lysine-coated black 96-well plates with clear bottom. At 16 to 24 h later, the medium is aspirated, and the cells are washed with 100 μL Krebs buffer [140 mM NaCl/4.7 mM KCl/2.5 mM CaCl2/1.2 mM MgCl2/11 mM HEPES/10 mM D-glucose, pH 7.4]; 50 μL Krebs buffer supplemented with various concentrations of UCPH-101 or TBOA is added to the wells, after which an additional 50 μL Krebs buffer supplemented with the FMP assay dye (1 mg/mL) is added to each well. The plate is incubated at 37°C in a humidified 5% CO2 incubator for 30 min and assayed in a reader measuring emission at 560 nm caused by excitation at 530 nm before and up to 1 min after addition of 33 μL Glu solution[1].

References:

[1]. Abrahamsen B, et al. Allosteric modulation of an excitatory amino acid transporter: the subtype-selective inhibitor UCPH-101 exerts sustained inhibition of EAAT1 through an intramonomeric site in the trimerization domain. J Neurosci. 2013 Jan 16;33(3):1068-87.