Dab substrate k176810e

Expression of SCARA5 in paraffin embedded lung cancer, adjacent paracancerous tissues, and xenograft tumor tissues in mice was detected by immunohistochemistry staining. The tissue was cut into 4 μm thick slices, transferred onto glass, and incubated at 65°C for at least 6 h. All the required reagents except antibody came from an Immunohistochemistry Kit (ZSGB-BIO, Beijing, China) and operation was performed according to the standard procedures. Slides were incubated at 4°C for 16–20 h with Ki67 (#16667, Abcam) and SCARA5 (#ab118894, Abcam) antibodies. Then tissues were stained with DAB substrate (K176810E, ZSGB-BIO, China) for 40–50 s. The nuclei were stained with hematoxylin for 5 s and covered with neutral resin. The image of IHC was observed by microscope. IHC scoring criteria: 0: The positive rate was less than 10%; 1: Positive rate was between 10 and 30%; 2: Positive rate was between 30 and 50%; 3: Positive rate was more than 50% added a description of Additional file 2: Table 2.

Immunohistochemistry (IHC) was used to detect expression of ZBTB16 in paraffin-embedded primary breast tumors and paired paracancerous tissues as well as xenograft tumor tissues in mice. Tissues were sliced into 4-μm-thick sections and baked overnight at 65 °C, washed with PBS and stained with an Immunohistochemistry Kit (ZSGB-BIO, Beijing, China) according to the manufacturer’s instructions. Slides were incubated at 4 °C overnight (16-20 h) with Ki67 (#16667, Abcam) and ZBTB16 (#ab104854, Abcam) antibodies. The next day, slides were stained with DAB substrate (K176810E, ZSGB-BIO, China) for 30 s and counterstained with hematoxylin for 3 s. Images were examined under a microscope.