Anti ibsp

5–7 μm thick sections of paraffin-embedded specimens were de-paraffinized in xylene, rehydrated in alcohol, and washed with phosphate-buffered saline (PBS) (Fisher Scientific). Antigen retrieval was performed at 95°C with sodium citrate (Sigma). Tissue sections were blocked for 1 hour with 5% bovine serum albumin (Sigma) and incubated with primary antibodies overnight. Primary antibodies used were as follows: anti-IBSP (rabbit polyclonal; 1:50 dilution; Abcam Cat. No. ab52128), antimetallothionein (mouse monoclonal; 1:00 dilution; Abcam Cat. No. ab12228), anti-CD45 (mouse monoclonal; 1:100 dilution; DAKO Cat. No. M0701), anti- MMP8 (rabbit monoclonal; 1:00 dilution; Abcam Cat. No. ab81286), and anti-αSMA (mouse monoclonal; 1:250 dilution; Sigma Aldrich Cat. No. A5228). Slides were incubated with FITC or TRITC-labeled secondary antibodies (1:500 dilution; Invitrogen, Carlsbad, CA) for 1 hour at room temperature and mounted with mounting media containing propidium iodide to visualize cell nuclei. Sections were viewed using a Nikon Eclipse E400 microscope and digital images were collected with QImaging camera and the NIS Elements BR 3.1 software.