Polink 1 hrp dab detection system one step polymer detection system

Tumor tissue were sectioned at the thickness of 5 μm and embedded in paraffin. To perform immunohistochemical staining, tissues were dewaxed and rehydrated in graded concentrations of xylene/alcohol. Antigen retrieval was performed in citrate buffer (pH 6.0) and heating at 121 °C. Sections were then blocked in goat serum (Boster, Wuhan, China) for 30 min at room temperature. Ki67 antibody (Bioss Antibodies, Inc., 1:200) was used to incubate the sections overnight at 4 °C. For TUNEL assay, Colorimetric TUNEL Apoptosis Assay Kit (Beyotime, Shanghai, China) was used to incubate the sections at 37 °C for 60 min. Following, Polink-1 HRP DAB Detection System One-step polymer detection system (ZSGB-BIO, Beijing, China) were added to the section and incubated for 20 min at room temperature. Hematoxylin was lastly used to stain the nucleus.

First, tumor tissue sections of mice were dried at 60°C for 1 h. Then, the sections were dewaxed in xylene and rehydrated in graded concentrations of alcohol. After that, the sections were treated by citrate buffer (pH 6.0) and autoclaved for 90 s at 121°C. After washing with PBS, sections were blocked with goat serum (Boster, Wuhan, China) for 30 min at room temperature and then incubated with Ki67 antibody (1:200, Bioss Antibodies, MA, U.S.A.) overnight at 4°C. Next, sections were washed by PBS and incubated using Polink-1 HRP DAB Detection System One-step polymer detection system (ZSGB-BIO, Beijing, China) for 20 min at room temperature. Finally, tissue sections were counterstained via hematoxylin.