Tissue sections were deparaffinized, dehydrared and then antigen retrieval was done in citric acid buffer (pH 6.0). The tissues were blocked with bovine serum albumin (BSA) for 1 hour before incubation overnight with the anti-IPO7 monoclonal antibody (1:100, sc365231, Santa Cruz) at 4 °C. The sections were then incubated with fluorescent secondary antibodies (1:200, A23210, Abbkine) for 1 hour. Then the tissues were washed 3 times in phosphate buffer saline (PBS), after which they were mounted in fluorescent mounting medium with DAPI (ZSGB-BIO). For cell immunofluorescence, the cells were fixed with 4% PFA at 4 °C, then permeabilized with 0.1% Triton X100 (BioFroxx), blocked by 3% BSA at 37 °C for 1 hour and were incubated with rabbit-anti-RUNX2 (YT5356, Immunoway) and mouse-anti-IPO7 (sc365231, Santa Cruz) antibodies overnight at 4 °C, then washed 3 times with PBS and incubated with fluorescent secondary antibodies for 1 hour (A23220, Abbkine; ANT034, Antgene). The fluorescence images were observed under fluorescence microscopy (Olympus 1 × 83, Japan).
Zhang Y., Zhang H., Yuan G., & Yang G. (2020). IPO7 Promotes Odontoblastic Differentiation of Mouse Dental Papilla Cells by Selectively Importing Odontogenic Transcription Factors.
+ Open protocol