Immunofluorescent staining of MEF1 cells using PA5-19589, anti-Gephyrin antibody. The cells were fixed with PFA (4%)for 10 minutes, permabilised TBS-T for 20 minutes and exposed to the primary antibody at a concentration of 1 ug/ml for 1 hour at room temp. A solution of BSA (1%), normal goat serum (10%) and glycine (0.3 M) was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody was a 448 fluorescence conjugated Goat anti-rabbit IgG (green) at a dilution of 1:1000. A WGA- 594 fluorescent conjugated stain was used to label plasma membranes (red) and the nuclei stain was DAPI (blue).