Immunofluorescence staining of embryos

1. Embryos were fixed in 4% paraformaldehyde for 15 min at RT.
2. Washed three times with PBS.
3. Permeabilized with 0.5% Triton X-100 in PBS for 15 min.
4. Blocking with 10% donkey serum for 1h at RT.
5. Incubated with the primary antibody in blocking solution for overnight at 4 ℃。
6. Embryos were washed with TBST for 6 times and 15min/time. 
7. Incubated with fluorescence-conjugated secondary antibodies (Invitrogen) for 2h at RT. 
8. Embryos were washed with TBST for 6 times and 15min/time.
9. Nuclei were counterstained with DAPI for 15 min.
10. Washed with TBST twice. 
11. Captured using the LSM 700.

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