GABA Release Quantification by ELISA

The procedure used to perform the assay was previously published [13 (link)]. The neurotransmitter levels were obtained and quantified using an ELISA (enzyme-linked immunosorbent assay) kit (#CEA900Ge; Cloud-Clone Corp.©, W. Fernhurst Dr., Katy, TX, USA) according to the manufacturer’s instructions. An ELX 800 IU automated Microplate Reader (Bio-Tek Instruments; Gene 5 Software V 3.02) was used to read the absorbance at 450 nm. A quadratic log-log curve fit was used to analyze the results. The supernatants (low and high K⁺ = before and after the GABA release) were analyzed for each well. In order to minimize the effect of the differences in the number of cells in the wells, the amount of GABA that had been produced per well was calculated based on the difference between the amount of GABA in a high K⁺ supernatant and the amount of GABA in a low K⁺ supernatant.