Ihc assay kit

Immunohistochemistry (IHC) staining of mouse tumor sections were performed according to standard protocols. In brief, paraffin-embedded sections were deparaffinized, rehydrated and subjected to antigen retrieval. After blocking with goat serum, sections were incubated with anti-E-cadherin (BD Biosciences) or anti-Vimentin antibody (Cell Signaling Technology) overnight at 4 °C, followed by secondary antibody incubation for 1 h in room temperature. DAB staining was performed with IHC assay kit (Maixin, Fuzhou, China).

IHC staining of paraffin-embedded human or mice tumor sections were performed according to standard protocols. Sections were deparaffinized, rehydrated, subjected to antigen retrieval, and blocked with 3% hydrogen dioxide and goat serum, followed by incubating in primary antibodies overnight at 4 °C. First antibodies used were listed as follows: anti-CD36 antibody (1:200, Abcam), anti-β-catenin (1:200, Genetex), anti-cmyc (1:200, GeneTex), anti-HK2 (1:500, ABclonal), anti-PKM2 (1:500, ABclonal), anti-GLUT1 (1:250, Abcam), anti-LDHA (1:250, GeneTex), anti-GPC4 (1:200, Proteintech), anti-GFP (1:500, Proteintech), anti-Ki-67 (for mouse, 1:800, Cell Signaling), anti-Ki-67 (for human, 1:800, Cell Signaling), anti-PCNA (1:1000, Proteintech). Next day, the sections were put in room temperature for 30 min to rewarm, followed by secondary antibody incubation for 1 h in room temperature and DAB staining was performed with IHC assay kit (Maixin, China). Counterstaining was carried with hematoxylin for 2 min. Images were taken with OLYMPUS DP22 microscope. Antibodies were shown in Supplementary Table 3.