Ki67 cell proliferation kit ihc

Nude mice on a BALB/c background (4–6 weeks old) were acquired from the Experimental Animal Center at Huazhong University of Science and adopted to engineer a tumor model in vivo. Huh7 cells stably transfected with miR-373-3p were adjusted to a cell concentration of 2 × 10⁷ mL-1. A total of 10 mice were injected subcutaneously in the right posterior axillary armpit with 0.1 ml cell suspension for each. In the next 5-week incubation, we monitored the survival rate, weight, and survival state of the mice. Since the 14th day, the tumor volume was gauged once every 4 days (volume = length to diameter × short diameter 2/2). Five weeks later, the mice were put to death, with their tumors harvested for weight measurement. The KI67 Cell Proliferation Kit (IHC) (Cat. no. E607235, Sangon Biotech (Shanghai) Co., Ltd.) was manipulated to examine Huh7 cell proliferation in line with the protocol. This animal experiment had received the green light from the Ethics Committee of The First Affiliated Hospital of Anhui Medical University and was implemented strictly in keeping with the guidelines of experimental animal care and use (NIH Publication No. 85-23201-1, National Institutes of Health, USA) [24 (link)].

The histological evaluation of tumor implanted tumors in nude mice were finished by combination of hematoxylin and eosin (H&E) staining and IHC (immunohistochemistry) assays. Briefly, tumor tissue sections (5 μm thick) were fixed with 4% neutral phosphate-buffered formalin at 4 °C, embedded with paraffin, and subjected to staining using the Hematoxylin-Eosin staining kit (#E607318; Sangon Biotech, Shanghai, China) according to the manufacturer’s instructions. The expression of Ki67 proteins in the tumor tissues were also detected by the Ki67 Cell Proliferation Kit (IHC) (#E607235; Sangon Biotech, Shanghai, China) following the producer’s instructions. The tissue slices were taken by a microscope (Olympus, Japan).