Vectastain elite abc hrp kit

Formalin-fixed paraffin-embedded (FFPE) tissue sections were cut at 5 μm and frozen tissues were cut at 6–8 μm for haematoxylin and eosin staining, and immunostaining. For tdTomato imaging, tissues were fixed in 10% neutral buffered formalin (Thermo Fisher Scientific) at 4°C overnight before embedding in OCT (Tissue-Tek, Sakura). Immunofluorescence staining on fresh frozen sections and immunohistochemistry (IHC) on FFPE tissue sections were performed as previously described (White et al., 2011 (link)). IHC was performed with antigen retrieval using antigen unmasking solution (citric acid based, Vector) for 30 min at 90°C. Antibodies were used as anti-S100 (1:200), anti-y-H2A.X (1:500), anti-Tenascin C (1:400), anti-SOX10 (1:200), anti-GFP (1:600), anti-Tyrp1 (1:200), anti-Trp2 (1:200), anti-F4/80 (1:400), anti-K5 (1:600), anti-CD45 (1:400), anti-Gr-1 (1:400), anti-Ki67 (1:200, #900-C01-B38 and 1:400, #14–5698), together with suitable Alexa-fluor or biotinylated secondary antibodies for IF and IHC. The IHC signal was amplified and detected using VECTASTAIN Elite ABC HRP Kit and AEC Peroxidase Substrate Kit followed by mounting using ImmunoHistoMount™ (Sigma). Prolong Gold antifade mountant with DAPI were used for IF staining.