Biotinylated goat anti mouse igg

The immunohistochemical studies were carried out using a standard streptavidin-biotin-peroxidase complex method demonstrated previously 18 (link). Briefly, tissue sections were deparaffinized and rehydrated. The endogenous peroxidase activity of the tissue was blocked using 3% hydrogen peroxide for 10 min. For antigen retrieval, slides were immersed and boiled in 10 mM citrate buffer (pH 6.0) for 15 min in a pressure cooker, and then non-specific binding was blocked by 5% normal goat serum for 10 min. The slides were incubated with anti-CHD1L (1:100 dilution; Abcam), a 1:200 dilution of monoclonal antibody against cluster of differentiation 82 (CD82), non-metastatic cells 4 (NME4), tumor necrosis factor (ligand) superfamily member 10 (TNFSF10) or methionyl aminopeptidase 2 (METAP2) at 4°C overnight in a moist chamber. The slides were sequentially incubated with biotinylated goat anti-mouse IgG (Santa Cruz Biotechnology) at a dilution of 1:100 and then allowed to react with the streptavidin-peroxidase conjugate, each for 30 min at room temperature. Staining was developed using the 3, 5-diaminobenzidine (DAB) Substrate Kit (Dako) according to the manufacturer's instructions, followed by Mayer hematoxylin counterstaining.

Specimens of human and rat were fixed, embedded, and sectioned into slides (4 µm thick). Sections were deparaffinized with xylene and rehydrated and blocked with 3% H₂O₂ to block endogenous expression. After antigen epitope repairing, sections were blocked with 5% normal goat serum for 20 min, the slides were incubated overnight at 4 °C to the mouse anti-PRL1 (Abcam, Cambridge, UK). Sections were then incubated with biotinylated goat anti-mouse IgG (Santa Cruz, Dallas, TX, USA) for 1 h. Samples were stained with 3,3’-diaminobenzidine (VECTOR Laboratories. Burlingame, CA, USA), followed by counterstaining with hematoxylin. Sections were imaged under a light microscope (Zeiss, Oberkochen, Germany).