Fig. 5
Lineage specificity of A7 MAb in bone marrow cultures for osteoclastogenesis. Bone marrow cells of 4-week-old SD male rats were cultured for 4 days in 96 multi-well plates at a concentration of (0.2 × 106 cells/well) as described in Materials and Methods. Unfixed cells were stained with A7 MAb or Kat-1 MAb as the primary antibody and further detected by ABC-AP kit (Vectastatin ®, Alkaline Phosphatase Standard, AK-5000) as described in Materials and Methods. Color development was performed using Vector Red (Vector® Red Alkaline Phosphatase substrate kit SK-5100) in the presence of 1 mM levamisole. Lower panels are the higher magnification view of the upper panels. a Specific staining of bone marrow stromal cells with A7 MAb (red stained cells). Arrowheads: Osteoclast-like MNCs expressing no A7 antigen. b Specific expression of Kat1 antigen in osteoclasts. Arrowheads: Osteoclast-like MNCs specifically stained with Kat-1 MAb (red). Bars, 50 µm
