Rabbit anti s100b

For immunohistochemistry, explants or free-floating tissue sections were rinsed in PBS. For myelin proteins, tissues were permeabilized 10% Triton X-100 in PBS (PBSTx) for 20 min and then rinsed in PBS. Tissues are blocked with 5% normal donkey serum (NDS) in PBSTx (1%) for 1h, and then incubated with primary antibodies overnight at room temperature. Following 3, 10 min washes in PBS, Alexa Fluor-labeled secondary antibodies (Jackson ImmunoResearch, West Grove, PA) were applied (1:800) 2h at room temperature, washed 3 times for 10 min in PBS, mounted on slides with water, dried and cover-slipped with Fluoromount G (Southern Biotech, Birmingham, AL). The following primary antibodies were used: Rat anti-PLP 1:100 (Yamamura et al., 1991 (link)), Rabbit anti-S100b 1:500 (Sigma, S2644), Rabbit anti-GFAP 1:1000 (Sigma, G9269), Goat anti-Iba1 1:400 (Abcam, ab5076), Guinea Pig anti-NG2 1:2000, Rabbit anti-NG2 1:2000, and Rabbit anti-PDGFRa 1:500 (Gifts from William Stallcup, Burnham Institute).

After treatment, cerebellar slices were rinsed twice with PBS and fixed in 4% paraformaldehyde in PBS for 20 min at room temperature. For immunohistochemistry, slices were rinsed in PBS and permeabilized in 1.5% or 10% (myelin proteins) Triton X-100 in PBS for 20 min. Slices were rinsed, blocked with 5% normal donkey serum (NDS) in PBS with 0.3% Triton X-100 for 1 h, and incubated with primary antibodies overnight at room temperature. Following 3 washes in PBS, Alexa Fluor-labeled secondary antibodies (Jackson ImmunoResearch, West Grove, PA) were applied (1:800) overnight at room temperature, washed 3 times in PBS and mounted in Fluoromount G (Southern Biotech, Birmingham, AL). The following primary antibodies were used: goat anti-Sox10 (Santa Cruz), rat anti-PLP (Yamamura, Konola, Wekerle, & Lees, 1991 (link)), mouse anti-AQP4 (Santa Cruz Biotechnology, Dallas, TX), rabbit anti-S100b (Sigma), rabbit anti-GST-pi (Enzo), mouse anti-Caspr (NeuroMAB), mouse anti-CC1 (Calbiochem), rabbit anti-GFAP (Sigma), rabbit anti-MOG (Abcam, Cambridge, United Kingdom), mouse anti-MBP (Covance, Princeton, NJ), chicken anti- Neurofilament-H (Neuromics, Minneapolis, MN), goat anti-Iba1 (Abcam).