Immunohistochemistry showed high expression of Bax in PCs

To confirm that PCs exposed to 3AP do contain Bax protein, we performed immunohistochemistry. Bax is a pro-apoptotic Bcl-2 family member, that plays a critical role in development and in tissue homeostasis, and its dysregulation can lead to neurodegenerative diseases and ischemic insults (Hara et al. 1996; Fan et al. 2001; Vogel 2002; Dorszewska et al. 2004; Jung et al. 2008; Didonna et al. 2012; Garcia et al. 2013). Cerebellar Purkinje cells and hippocampal neurons have the highest levels of Bax (Hara et al. 1996; Vogel 2002; Casoni et al. 2017). Mouse and Rabbit specific horseradish peroxidase (HRP)/3, 3' Diaminobenzidine (DAB) Detection IHC Kit ab64264 is a complete kit to detect a primary antibody raised in either mouse or rabbit, using HRP-labeled-streptavidin and a biotinylated anti-rabbit/anti-mouse secondary antibody. It is supplied with a full protocol and with peroxidase and protein blocking reagents, biotinylated secondary antibody, streptavidin-HRP, and DAB substrate. A sample is incubated with an unconjugated primary antibody specific to the target antigen, a biotinylated secondary antibody then binds to the primary antibody, HRP-labeled streptavidin then binds to the secondary antibody. Visualization is based on enzymatic conversion of a chromogenic substrate DAB into a colored brown precipitate by HRP at the sites of antigen localization, which can then be viewed using brightfield microscopy.