After blocking in 5% NDS, 0.5% Triton X-100 in TBS at 4°C overnight, samples were incubated at 4°C overnight with the primary NICD antibody diluted in blocking buffer (1:400). After washes, embryos ...

Triton X-100, SDS at a final concentration of 0.25%, 0.5% and 1%; and an oxidizing agent-H 2 O 2 at a final concentration of 0.5%, 1%, 1.5% and 2%. The incubated enzyme (without inhibitors, detergents ...

Free-floating sections were rinsed in PBS and incubated with 10% normal donkey antiserum (NDS) and 0.2% Triton X-100 in PBS for 60 min to reduce nonspecific staining and increase antibody penetration.