Immunohistochemical Profiling of Tumor Tissue

IHC staining of tumor tissue using AEG-1/MTDH-specific rabbit polyclonal antibody (#13860-1-AP), MAP-LC3 (CST-12741), TGF-β1 (CST-3711), E-cadherin (BS-1097) and N-cadherin (BS-2224) antibodies is performed as described previously [39 (link)]. Briefly, human tumor samples are subjected to deparaffinization, rehydration, and antigen retrieval before the staining procedures are performed. The tissue slides are blocked with 2.5% normal horse serum for 10 minutes. Then, tissue slides are incubated with AEG-1/MTDH-specific rabbit polyclonal antibody, MAP-LC3, TGF-β1, E-cadherin and N-cadherin antibody (dilution 1:50) overnight at 4°C. After the tissue slides are washed, they are incubated with anti-mouse IgG HRP and anti-rabbit IgG HRP secondary antibody for 10 minutes. The slides are stained with 3, 3′-diaminobenzidine (DAB) (Vector Laboratories), counterstained with hematoxylin (Vector Laboratories), dehydrated, treated with xylene, and mounted. All slides are examined and representative pictures are taken using an Olympus B × 41 microscope (Olympus America, Melville, NY) [40 (link)].