Dab detection kit streptavidin biotin

The first molars were dissected from the E14.5 d, E16.5 d, E18.5 d, P.N.4 d, P.N.7 d, P.N.10 d, and P.N.15 d rats (Figures 1, 2) and the E16.5d p75NTR^ExIII−/− and p75NTR^ExIII+/+ mice (Figure 4), fixed in 4% paraformaldehyde, decalcified with 10% EDTA, and embedded in paraffin. The 6-μm sections of tissue specimens were obtained for H.E. and immunohistochemistry staining. The primary antibodies were used in this study are as follows: rabbit anti-rat p75NTR (1:1,500; Abcam, Cambridge, MA, United States, ab245134, monoclonal), rabbit anti-rat BMAL1 (1:1,000; Abcam, Cambridge, MA, United States, ab230822, monoclonal), rabbit anti-rat CLOCK (1:1,000; Abcam, Cambridge, MA, United States, ab3517, polyclonal), rabbit anti-rat PER1 (1:500; Bioss, Beijing, China,bs-2350R, polyclonal), rabbit anti-rat CRY1 (1:500; Bioss, Beijing, China bs-11441R, polyclonal), rabbit anti-rat ALP (1:500; Bioss, Beijing, China, bs-2928R, polyclonal), rabbit anti-rat COL1 (1:500; Bioss, Beijing, China, bs-0578R, polyclonal). These specimens were treated with the DAB Detection Kit Streptavidin-Biotin (ZSGB-BIO, Beijing, China) and Hematoxylin and Eosin Staining Kit (Beyotime, Shanghai, China) according to the manufacturer’s protocols, followed by visualization under phase-contrast microscopy.

Embryonic maxillofacial processes were dissected from mice at E13.5 d, E15.5 d and E18.5 d. The tissues were fixed in 4% paraformaldehyde and sectioned at 6 μm for immunostaining. The following primary antibodies were used: rabbit anti‐mouse p75NTR (ab8847, 1:1500; Abcam) and rabbit anti‐mouse Mage‐D1 (sc‐28243, 1:1500; Santa Cruz Biotechnology). Then, the specimens were treated with the DAB Detection Kit Streptavidin‐Biotin (ZSGB) according to the manufacturer's protocols, followed by visualization under a phase‐contrast microscope.