Immunofluorescence staining of undifferentiated SH-SY5Y cells (Human neuroblastoma, ATCC CRL-2266) (left) and differentiated SH-SY5Y cells (right). Undifferentiated cells were seeded in a collagen coated 384-well imaging plate at ~ 8,000 cells per well. After overnight incubation, cells were stained using the methanol fix/perm protocol and the mouse anti-GRIP antibody. Differentiated cells were seeded in a 96-well, collagen coated imaging plate at ~ 5,000 cells per well. Cells were incubated with 50 mM ATRA (Sigma-Aldrich) for 5 days, followed by 50 ng/ml BDNF (Sigma-Aldrich) for 5 days. Differentiated cells were fixed and stained using the methanol fix/perm protocol, and the mouse anti-GRIP antibody. The second step reagent in both cases was Alexa Fluor® 488 goat anti-mouse Ig (Invitrogen). The images were taken on a BD Pathway™ 855 or 435 Bioimager using a 20x objective. This antibody also stained undifferentiated SK-N-SH (Human neuroblastoma, ATCC HTB-11), C6 (Rat glioma, ATCC CCL-107), U-87 MG (Human glioblastoma cells, ATCC HTB-14) and U-373 cells (Human glioblastoma cells, ATCC HTB-17, discontinued) using both the Triton-X 100 and methanol fix/perm protocols.