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Immunohistochemistry (IHC) on human samples

SG Simone Di Giacomo
MS Manuela Sollazzo
DB Dario de Biase
MR Moira Ragazzi
PB Paola Bellosta
AP Annalisa Pession
DG Daniela Grifoni
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IHC staining of tumour samples was carried out on 3μm formalin-fixed paraffin-embedded (FFPE) sections. YAP staining (rabbit polyclonal antibody, Cell Signaling Technology #4912, 1:150)61 and HUGL-1 staining (rabbit polyclonal antibody, D. Strand, 1:550)37 were performed on consecutive slices using UltraVision Kit (Thermo Scientific) and the DAB Chromogen System (Dako). c-MYC staining (mouse monoclonal antibody 9E10, DSHB, 1:50)62 and Cleaved-Caspase-3 staining (rabbit polyclonal antibody, #9961 Cell Signaling Technology, 1:250)63 were performed on the same slice using the EnVision G/2 Doublestain System (Dako). Cell nuclei were counterstained with Mayer’s Hematoxylin. Slides were scanned using an Aperio Scanscope CS System by Leica Biosystems and original digital frames were used to compose the figure panels. Samples were from the Bellaria Hospital archives (Bologna, Italy), where informed consent was obtained prior to sample banking. Institutional Review Board approval (Nr. 20OS11) was obtained at the Bologna AUSL (Azienda Unità Sanitaria Locale). All experiments were performed in accordance with relevant guidelines and regulations.

Copyright and License information: The Author(s) ©2017
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