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Table 1 Details of the immunohistochemistry methodology employed

From: An in vitrostudy investigating the survival and phenotype of mesenchymal stem cells following injection into nucleus pulposus tissue

Target Antigen retrieval Blocking step and primary antibody Secondary antibody
Aggrecan 0.1% w/v hyaluronidase in Tris-buffered saline (Sigma, Poole, UK), 30 minutes at 37°C 20% v/v rabbit serum, and mouse monoclonal aggrecan 1° (1:25 dilution; AbCam, Cambridge, UK) Biotinylated rabbit anti-mouse antiserum (1:400; Dako, Ely, Cambridgeshire, UK)
Sox-9 None required 20% v/v swine serum, and rabbit polyclonal Sox-9 1° (1:100 dilution; SantaCruz, Heidelburg, Germany) Biotinylated swine anti-rabbit antiserum (1:400; SantaCruz)
Type I collagen 0.01% hyaluronidase (Sigma), 0.02% trypsin (Sigma) w/v in Tris-buffered saline 20% v/v rabbit serum, and mouse monoclonal type I collagen 1° (1:250 dilution; ICN, Basingstoke, UK), Biotinylated rabbit anti-mouse antiserum (1:400; Dako)
Type II collagen 0.1% w/v hyaluronidase in Tris-buffered saline (Sigma), 30 minutes at 37°C 20% v/v rabbit serum, and mouse monoclonal type II collagen 1° (1:100 dilution; MP Biomedicals, Illkirch, France) Biotinylated rabbit anti-mouse antiserum (1:400; Dako)
Type X collagen 1 mg/ml hyaluronidase, 0.25 U/ml chondrotinase in Tris-buffered saline, 1 hour at 37°C; followed by 0.1% protease in Tris-buffered saline, 10 minutes at 37°C 25% v/v goat serum, and mouse monoclonal type X collagen 1° (1:200 dilution; AbCam) Biotinylated goat anti-mouse antiserum (1:100; SantaCruz)
Caspase 3 None required 20% v/v donkey serum, and goat polyclonal caspase 3 1° (1:500 dilution; SantaCruz) Biotinylated donkey anti-goat antiserum (1:300; SantaCruz)