Immunocytochemical Staining Method
1. Place the cell specimen on a glass slide;
2. Fixation: It is recommended to fix with 95% ethanol or 10% neutral buffered formalin for 10 minutes;
3. Rinse slightly, add 1% Triton X-100 reagent to the slide, incubate at room temperature for 10 minutes, and wash with PBS 3×3 minutes;
4. Perform appropriate antigen retrieval on the cells according to the requirements of the primary antibody;
5. If necessary, add peroxidase blocking reagent to each slide, incubate at room temperature for 10 minutes, and wash with PBS 3×3 minutes;
6. Remove PBS, add the primary antibody to the slide, incubate at room temperature for 60 minutes or at 4°C overnight,Wash with PBS 3×3 minutes;
7、Remove PBS, add the detection kit to the slide (operate according to the kit instructions),Wash with PBS 3×3 minutes;
8、Remove PBS, select the corresponding chromogenic system according to the detection kit, and finally mount with neutral resin.
Note:A better method is to gather the cells by centrifugation or direct collection, then wrap them with egg white or agar to form a cell block for fixation, dehydration, embedding, sectioning, and then follow the immunohistochemical staining steps.