Rabbit anti sox10

Dechorionated embryos and larvae were fixed with 4% PFA for 1 h at room temperature (RT), washed in PBSTX (1% Triton X-100, 1× PBS) for 5 min, followed by a 5 min wash with DWTx (1% Triton X-100 in DH₂O), then permeabilized in acetone at RT for 5 min and at −20°C for 10 min, followed by a 5 min wash with PBSTx. Embryos were then blocked in 5% goat serum/PBSTx for 1 h, incubated in primary antibody with 5% goat serum/PBSTx for 1 h at RT and overnight at 4°C. Embryos were washed extensively with PBSTx at RT and incubated in secondary antibody overnight at 4°C. After antibody incubation, embryos were washed extensively with PBSTx and stored in PBS at 4°C until imaging. The following antibodies were used: rabbit anti-Sox10 (1:5000) (Binari et al. 2013 (link)), Alexa Fluor 647 goat anti-rabbit IgG(H+L) (1:1000; ThermoFisher). Embryos were mounted in glass-bottomed Petri dishes for imaging as described above.