Goat anti amylase

Tissue specimen processing, sectioning, and H&E staining were performed using standard protocols. Immunohistochemistry was performed using the VectaStain Elite ABC kit (Vector Laboratories) according to the manufacturer's protocol. The antibodies used were mouse anti-MUC5AC (1:500; ThermoFisher), mouse anti-Ki67 (1:100; BD Pharmingen), rat anti-Ck19 (1:750; University of Iowa), and goat anti-amylase (1:100; Santa Cruz Biotechnology). The sections were counterstained with hematoxylin or Alcian blue/nuclear fast red using the NovaUltra Alcian blue stain kit (IHC World) according to the manufacturer's instructions. For Ck19 and amylase staining, the sections were stained using anti-goat Alexa 488 (1:200; Invitrogen) and anti-rat Alexa 594 (1:200; Invitrogen) and counterstained with DAPI. Pictures were taken using a Leica microscope and/or with a NanoZoomer 2.0-RS slide scanner (Hamamatsu). Analysis of the PanIN and mucinous cystic lesion areas and Ki67 staining was performed using ImageJ. To simulate the size criterion used to diagnose IPMNs in humans, we also used a size criterion (diameter ≥280) to call cystic lesions/IPMNs. This size was based on the ability to distinguish large cystic lesions from PanIN lesions. Further classification of these lesions was performed based on their lining using H&E and Muc5ac staining.