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2.12. Immunohistochemical staining

XZ Xiaozhu Zeng
MZ Maoxi Zhong
YY Yumeng Yang
ZW Zhi Wang
YZ Yuxi Zhu
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Tumours allografts were removed and fixed in 4% paraformaldehyde. 4 μm thick paraffin sections were prepared from allografts for immunohistochemistry (IHC) staining. The slides were then deparaffinized with xylene, and rehydrated by using graded ethanol. Antigen retrieval was performed by using microwave heating for 20 minutes in sodium citrate‐hydrochloric acid buffer. Next, the sections were incubated at 4°C overnight with primary antibody: the anti‐RCC1 rabbit antibody (1:500, ABclonal, China), anti‐p27kip1 rabbit antibody (1:500, ABclonal, China), anti‐CDK4 rabbit antibody (1:500, ABclonal, China) and anti‐PD‐L1 rabbit antibody (1:500, ABclonal, China). Biotinylated goat anti‐mouse or goat anti‐rabbit antibody was used as a secondary antibody for 20 minutes at room temperature, followed by colour development with DAB. Ultimately, the slides were counterstained with haematoxylin. A light microscope was used to observe and capture the images of histopathological changes.

Copyright and License information:  ©2021 The Authors. published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd.
This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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