Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

Immunohistochemical staining of cleaved caspase-3

SO Shouichi Okamoto
HE Hiroki Ebana
MK Masatoshi Kurihara
KM Keiko Mitani
EK Etsuko Kobayashi
TH Takuo Hayashi
YS Yasuhito Sekimoto
KN Koichi Nishino
MO Mizuto Otsuji
TK Toshio Kumasaka
KT Kazuhisa Takahashi
KS Kuniaki Seyama
request Request a Protocol
ask Ask a question
Favorite

Lung tissues resected during surgery for SP were fixed in 10% buffered formalin and embedded in paraffin. After deparaffination of lung tissue specimens, antigen retrieval was sequentially conducted by autoclave in 10 mM Citrate buffer (pH 6.0) for 15 min at 105 ℃. Samples were incubated with anti-rabbit cleaved caspase-3 antibody (dilution 1:200; #9664, Cell Signaling Technology, Danvers, Massachusetts, USA) for 1 h at room temperature following incubation with Histofine Simple Stain MAX-PO (MULTI) (#424152; Nichirei, Tokyo, Japan) for 30 min at room temperature. Peroxidase staining was visualized with 3,3-diaminobenzidine and hematoxylin was used for counter staining. At least 300 PMCs were counted in each sample and the percentage of PMCs stained with anti-cleaved caspase-3 antibody was calculated in 3 unrelated samples per group.

Copyright and License information: The Author(s) ©2021
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

post Post a Question
0 Q&A