Rabbit anti vdr

Immunohistochemical analysis involving animals followed protocols approved by the Institutional Animal Care and Use Committee of Seoul National University (SNU-180207-3). Six-week-old Sprague Dawley rats were sacrificed by CO₂ asphyxiation. Mandibular molars, including the surrounding tissues, were fixed in 4% paraformaldehyde in phosphate buffer (pH 7.4) at 4°C overnight. Decalcified tissues in a solution of 10% ethylenediaminetetraacetic acid (pH 7.4) were embedded in paraffin, and then processed for the immunohistochemical examinations. The deparaffinized sections were treated to retrieve antigens in a solution of citrate buffer (pH 6.0) at 60°C overnight. Expression of VDR was detected using an ABC kit (Vector Labs, Burlingame, CA, USA) and a DAB kit containing nickel chloride to enhance the contrast of the immunoreactivity (Vector Labs) after treating tissues with rabbit anti-VDR (1:50, Santa Cruz, Santa Cruz, CA, USA) as the primary antibody and a biotin-labeled goat anti-mouse IgG (1:200, Vector Labs) as the secondary antibody. The negative control slides were treated similarly, except that the reaction with the primary antibody was omitted.