Ab26116

Inhibitory human neurons were fixed for 15 minutes in 4% paraformaldehyde in phosphate buffered saline (PBS) and permeabilized using 0.1% Triton X-100 in PBS for 10 minutes at room temperature. Cells were then incubated in blocking buffer (4% bovine serum albumin (BSA) with 1% normal goat serum in PBS) for 1 hour at room temperature and then incubated with primary antibodies diluted in blocking buffer for 1 hour at room temperature, washed with PBS three times, and subsequently incubated in secondary antibodies for 1 hour at room temperature. Confocal imaging analysis was performed using a Zeiss LSM700. Primary Antibodies used include: mouse anti Oct4 (Millipore Sigma MAB4401, 1:2000), mouse anti Tra-1-60 (Millipore Sigma MAB4360, 1:1000), mouse anti MAP2 (Sigma-Aldrich M1406, 1:500), rabbit anti MAP2 (Sigma-Aldrich M3696, 1:500), chicken anti MAP2 (Millipore AB5543, 1:1000), rabbit anti Synapsin (e028, 1:3000), rabbit anti VGAT (Millipore Sigma AB5062P, 1:2000), mouse anti Gad-67 (Abcam ab26116, 1:500), mouse anti β3 Tubulin (BioLegend 801201, 1:2000), mouse anti Calbindin (Abcam ab82812, 1:500), guinea pig anti Parvalbumin (Swant GP72, 1:2500), rat anti Somatostatin (Millipore MAB354, 1:50).