Location of SIBLING proteins in mineralized dentin.
An immunocytochemistry analysis using one of two primary antibodies, polyclonal antidentin sialophosphoprotein (Abcam, Cambridge, MA, USA) which is specific to dentin sialoprotein (DSP) or polyclonal antidentinal matrix protein (DMP)-1 (Sigma-Aldrich, St Louis, MO, USA) and visualized using peroxidase (brown color in (a) and (c)) or Alexa Fluor 594 (red color in (b) and (d)) coupled streptavidin under fluorescence microscopy revealed the localization patterns of DSP and DMP-1. DSP and DMP-1 are strongly expressed in the odontoblastic layer (arrow) and with less intensity in the predentin area (double arrow) and in the dentin mineralization front (head arrow). (a) and (b) immunopositive staining for DSP in the mineralized dentin. (c) and (d) immunolocalization of DMP-1. These non-collagenous proteins interact with collagen fibrils and control initiation and growth of apatite crystals on dentinal matrix. Mineralized dentin counterstained with hematoxylin (a and c) and Hoechst blue 33342 (b and d). Bar corresponds to 100 μ
m. Courtesy of Baldion et al. [29