Fig 1: Presence of tuft cell morphology markers in ChAT-immunoreactive tuft cells. Cytokeratin 18 (CK18) immunoreactivity is present in many, if not all epithelial cells from the small intestine (A), peribiliary glands (B), and pancreatic ducts (C). Hence, double-immunofluorescence analysis of ChAT with CK18 revealed complete co-expression in human small intestine (Aa–c), peribiliary glands (Ba–c, arrow points to ChAT/CK18 co-immunoreactive cell), and pancreatic ducts (Ca–c). Villin (VIL) immunoreactivity is present at the apical brush border of almost all intestinal epithelial cells (D). Local accumulations indicate location of tuft cells (arrow in D), which is confirmed by double labeling of ChAT with VIL (Da–c, arrowhead points to VIL accumulation in tuft cell brush border). VIL immunoreactivity is also present in solitary cells in extrahepatic peribiliary glands (E) and in pancreatic ducts (F). Again, ChAT immunoreactivity is present in VIL immunoreactive cells (Ea–c, and Fa–c); arrowheads demarcate VIL immunoreactivity in apical tuft). Advillin (AVIL) immunoreactivity is restricted to solitary cells in the epithelial lining of the small intestine (G, asterisks), peribiliary glands (H), and pancreatic ducts (I). In all these locations AVIL co-localized to ChAT (Ga–c, Ha–c, Ia–c). The single antibody labeling (a,b) is shown in greyscale, the composite (c) in green (ChAT) and in red (CK18, VIL, AVIL) color coding, respectively. For all double-immunofluorescent analyses of ChAT, the goat-anti-ChAT antiserum was used. The bar in A equals 20 µm and applies to all bright-field images. The bar in Aa equals 10 µm and applies to all images from immunofluorescence analysis.
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