Alexa fluor 488 labeled goat anti mouse igg

The gastric cancer tissue block was embedded in paraffin and continuously cut into 3 µm sections and placed on a glass slide. The slices were baked at 72 °C for 1 h, dewaxed with xylene, and dehydrated with gradient alcohol. After rinsing five times with phosphate-buffered saline (PBS), 2 min each time (1‰ Tween 20 is added to PBS), then after high-pressure repair, rinse again with PBS five times. The processed sections were immersed in a 3% hydrogen peroxide solution, incubated at room temperature for 10 min, and washed with distilled water and PBS. Next, CD8 mouse-derived primary antibody (Santa Cruz, sc-70794, USA; 1:100) was added to the slices in equal proportions, incubated at 37 °C for 1 h, and rinsed with PBS three times for 5 min each time. Alexa Fluor 488-labeled goat anti-mouse IgG (Jackson, 115-545-003, USA; 1:1000) was added in equal proportions of secondary antibody mixture. After incubating at 37 °C for 25 min, the cells were rinsed with PBS three times for 5 min each time, allowed to dry and mounted with 4’,6-diamidino-2-phenylindole, dihydrochloride (Invitrogen, S36942, USA). An AXIO Scan. Z1 scanner was used to scan. Finally, the percentage of positive cells was counted, and the results were confirmed by two pathologists.