Ab189776

After being fixed and decalcified, the blocks were processed into routine slices and then underwent dewaxing with xylene and hydration with gradient alcohol. The slices were applied with 3% H2O2, followed by 15-min resting to blockade endogenous peroxidase activities remaining in tissues, and then were washed with PBS triply. The ccRCC slices were applied with the citrate solution (10 mM) and heated in microwave oven for 20 min for antigen retrieval. The ccRCC slices were subsequently applied with 5% serum of goat for 10 min, for the sake of blocking the nonspecific binding. After applying with the primary antibodies of CD8 (1/200, ab4055, Abcam, UK), CD45RO (5 μg/ml, ab23, Abcam), HMGB3 (1/100, ab75782, Abcam), IKBKE (10 μg/ml, ab7891, Abcam), PLTP (1/50, ab189776, Abcam), PTGES (10 μg/ml, ab62050, Abcam) and CD36(1:100 ab133625, Abcam), the slices were preserved at 37°C for 0.5 hour and rinsed with PBS. After applying with the secondary antibody (18653, Cell Signaling Technologies, USA) labelled horseradish peroxidase, the slices were preserved at 37°C for 0.5 hour and rinsed with PBS once again, stained utilizing the diaminobenzidine (DAB) (91-95-2, Sigma-Aldrich Chemical Company, USA) for 10 min, rinsed with running purified water, stained with hematoxylin, added with neutral resins and eventually observed utilizing the microscope and the pictures were taken.