Table 1 Antibodies select for IHC, antigen retrieval procedure, optimized dilution and incubation period
Antibody | Antigen Retrieval | Incubation | Reference / Brand |
|---|---|---|---|
Collagen type I | 10 mM citrate buffer, pH=6.0, 1’ MW 1,5 U/mL hyalurinodase (Sigma-Aldrich), 45’ 37ºC | Dilution: 1:100 Incubation overnight | 600-401-103-0.1 / Rockland Immunochemicals, Inc. Limerick, PA |
Fibronectin | 10 mM citrate buffer (pH=6.0), 1’ MW 1,5 U/mL hyalurinodase (Sigma-Aldrich), 45’ 37ºC | Dilution: 1:50 Incubation overnight | SC-8422 / Santa Cruz Biotechnology |
Alpha Smooth Muscle Actin | 10 mM citrate buffer (pH=6.0), 1’ MW 1,5 U/mL hyalurinodase (Sigma-Aldrich), 45’ 37ºC | Dilution: 1:400 Incubation overnight | SC-53015 / Santa Cruz Biotechnology |
Matrix Metalloproteinase 12 | 10 mM citrate buffer (pH=6.0), 1’ MW 1,5 U/mL hyalurinodase (Sigma-Aldrich), 45’ 37ºC | Dilution: 1:400 Incubation overnight | SC-390863 / Santa Cruz Biotechnology |
Collagen type II | 10 mM citrate buffer (pH=6.0), 1’ MW 20 μg/mL proteinase K (Sigma-Aldrich), 15’ 37ºC | Dilution: 1:100 Incubation overnight | SC-52658 / Santa Cruz Biotechnology |
CD68 | Tris-EDTA buffer (pH=9.0), 1’ MW | Dilution: 1:100 Incubation 30’ | PG-M1 / Dako |