Rabbit anti fibronectin antibody

DPSCs cultured on the hydrogels and dishes were rinsed with PBS, fixed in 4% paraformaldehyde for 15 min at room temperature (RT). Before immunofluorescence staining, fixed DPSCs were blocked using 3% BSA supplemented with 0.3% Triton X-100 for 30 min at RT. Then, cells were incubated for 2 h with primary antibody and washed twice with PBS. After secondary antibodies staining for 1 h at RT, nuclei were stained as the manufacturer’s instruction (Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA). Cell imaging was performed with a laser scanning confocal microscope (LSM710; Zeiss, Jena, Germany).
The primary and secondary antibodies in this examination mainly included as follows: rabbit anti-Collagen I antibody (Abcam, MA, USA), rabbit anti-Fibronectin antibody (Abcam, MA, USA), mouse anti-N Cadherin antibody (Abcam, MA, USA), mouse anti-E Cadherin antibody (Cell Signaling Technology, USA), Alexa Fluor 488 Donkey anti-Mouse IgG (Invitrogen, Eugene, OR, USA) and Alexa Fluor Plus 594 Donkey anti-Rabbit IgG (Invitrogen, Eugene, OR, USA).

Tissue sections (5 μm) were made as described above. After deparaffinizing, rehydrating, antigen retrieval, and blocking, the sections were incubated overnight at 4°C with rabbit anti-collagen I antibody (Abcam), rabbit anti-collagen III antibody (Abcam), rabbit anti-fibronectin antibody (Abcam), and rabbit anti-alpha smooth muscle actin (α-SMA) antibody (Abcam), respectively. On the next day, samples were washed with phosphate-buffered saline (PBS) 3 times for 5 min each time, before incubation with goat anti-rabbit secondary antibody labeled with horseradish peroxidase (KeyGEN Biotechnology) for 30 min. Diaminobenzidine (DAB) (KeyGEN Biotechnology) was used for coloring. Immunohistochemical assay of these outcomes was performed using Image Pro Plus software.