Rabbit anti tia1 antibody

The core components of SG, T-cell intracellular antigen-1 (TIA1), and GTPase-activating protein-binding protein 1 (G3BP1) are considered direct indicators of SG formation. Therefore, we used a double-labeled immunofluorescence (G3BP1 and TIA1) assay to observe SG generation in the cortex, PC12 cells, and primary cortical neurons. For immunofluorescent staining, frozen sections (10 μm thick) of the brain and cells (PC12 cells and primary cortical neurons) mounted on cover glass were fixed with 4% paraformaldehyde (PFA) for 20 min, permeabilized in 0.3% Triton X-100 solution for 10 min, blocked with blocking solution (5% BSA and 10% goat serum) for 1 h and finally incubated with primary antibodies (rabbit anti-TIA1 antibody, 1:1000, Abcam; mouse anti-G3BP1 antibody, 1:1000, Abcam, Cambridge, United Kingdom) and secondary antibodies (Alexa Fluor488-conjugated goat anti-rabbit antibody, 1:500, Abcam; Alexa Fluor555-conjugated goat anti-mouse antibody,1:500, Abcam) in the dark. DAPI staining for 10 min was used to label nuclei. Images were acquired using a confocal laser scanning microscope (LSM 800, Zeiss, Oberkochen, Germany). Each experiment was replicated three times and data are presented as the mean ± SEM.