1. Dewax and rehydrate FFPE sections in xylene and decreasing concentrations of IMS.
2. Wash sections in dH2O for 5 min.
3. Heat-induced antigen retrieval (if required)
4. Wash sections in dH2O for 5 min, followed by TBS-T 0.1% for 2x 5 min.
5. Blocking with normal goat serum in TBS-Triton X 0.25% for 1 h.
6. Overnight primary antibody in TBS-Triton X 0.1% incubation at 4 C.
7. Wash sections in PBS (no detergents) for 2x 5 min.
8. Incubate with 0.3% Sudan Black
9. Wash sections in PBS for 5 min.
10. Incubate with TrueVIEW for 5 min.
11. Wash sections in PBS for 5 min.
12. Incubate with fluorescent secondary antibodies in PBS for 1 h at room temp.
13. Wash sections in PBS for 2x 5 min.
14. Coverslip using VECTASHIELD Vibrance * and allow to set for 2 h.
*VECTASHIELD Vibrance included in TrueVIEW Kits, but may also be purchased separately