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URL: https://dx.doi.org/10.17504/protocols.io.bkqykvxw

Authors: Lora Starrs, Debra Brown, Sarah Popp, Andrew Ziolkowski, Charmaine Simeonovic

Summary: Paraffin sections (4 μm thickness) of formalin-fixed human pancreases and isolated human islets were treated with 0.05% pronase for antigen retrieval. HS and Col18 HSPG core proteins were detected immunohistochemically using 10E4 anti-HS (US Biological/Amsbio) and anti-Col18 (Santa Cruz), respectively, with horseradish peroxidase-conjugated rabbit anti-mouse Ig (Dako). Background staining was checked using the corresponding isotype control Ig instead of the primary antibody. 3-amino-9-ethylcarbazole (AEC) was used as the chromogen. For morphometry, stained sections were imaged using a light microscope with attached camera (Olympus BX41). Image J software with color deconvolution plugin was used for the quantitative analysis of the % of islet area stained

Affiliations: The John Curtin School of Medical Research, The Australian National University, The John Curtin School of Medical Research, The Australian National University, The John Curtin School of Medical Research, The Australian National University, The John Curtin School of Medical Research, The Australian National University, The John Curtin School of Medical Research, The Australian National University

External URL: https://doi.org/10.1371/journal.pone.0191360

Version: 2

Publication Date: 2020

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