Rabbit anti ocn primary antibody

At 6 weeks after cranial defect treatment, the samples were collected for hematoxylin and eosin (H&E) staining. The samples were fixed by 10% formalin for 24 hours. After that, 10% EDTA‐2Na+ 1% NaOH were utilized to decalcify for 24 hours, and then were embedded in paraffin and sectioned (8 μm thicknesses). Samples were also collected for immunohistochemical (IHC) stainings to analyses the expression of the OCN gene. Briefly, sodium citrate buffer was used to retrieve antigen for 20 minutes at 95°C, nonspecific binding was suppressed with for 1 hour at room temperature. Samples were incubated with rabbit anti‐OCN primary antibody (1:200) (Abcam ab93876, Cambridge, Massachusetts) overnight at 4°C, then incubated with horse anti‐rabbit biotinylated secondary antibodies (Vector Laboratories, Burlingame, California) 30 minutes. Samples were incubated with horseradish peroxidase‐conjugated streptavidin (Vector Laboratories) and treated with the Vector NovaRED peroxidase substrate (Vector Laboratories). After IHC, samples were then subsequently counterstained with hematoxylin, dehydrated, mounted, cover slipped. The images were captured by the Olympus CKX41 microscope.