Alexa fluor 633 goat anti chicken

Immunofluorescence was performed as previously reported (Ismail et al. 2017 (link)) on hippocampal primary neurons after administration of the treatments indicated above. Single and double immunofluorescence was performed with the following primary antibodies: chicken anti-MAP2 (1:10 000 dilution, Abcam, ab5392) and mouse anti-PSD95 (1:1000 dilution, Abcam, ab2723). Cells were then incubated for 1 h at room temperature at a dilution of 1:1000 with secondary antibodies; Alexa fluor 633 goat antichicken and Alexa flour 488 goat antimouse (Invitrogen, A11035 and A11055, respectively). Identification of the nuclei of cell bodies was performed using 4, 6 diamino-2-phenylindole (DAPI; Sigma) and the F-actin was stained by Alexa Fluor 488-Phalloidin (1:1000; Life Technologies). Omission of the primary antibody was done as a control staining. Coverslips were finally mounted onto glass slides using fluorescence mounting medium (DAKO Cytomation, Glostrup, Denmark). Imaging was performed with a Zeiss (LSM 510 META) confocal laser scanning system using a 488 nm Argon laser, 633 nm Helium–neon laser and UV 405 nm. The fluorescence of DAPI, Alexa 488 and Alexa 633 was recorded through separate channels.