Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

Immunofluorescence assay

YZ Yi Zhao
JY Juan Yang
JL Jiarui Liu
YC Yiqing Cai
YH Yang Han
SH Shunfeng Hu
SR Shuai Ren
XZ Xiangxiang Zhou
XW Xin Wang
ask Ask a question
Favorite

DLBCL cells treated with designed concentrations of CFI-400945 or DMSO for 24 h were fixed with 4% paraformaldehyde at room temperature for 15 min. Thereafter, cells were permeabilized with 0.4% Triton X-100 for 10 min. The slides were then incubated in blocking solution at room temperature for 1 h, followed by incubation with primary antibodies against YAP and γ-H2AX overnight at 4 °C and then with Alexa Fluor 488-labeled secondary antibody (Abbkine, Beijing, China). Nuclear was stained with DAPI. Microfilaments were stained with phalloidin (Abcam, Cambridge, MA, USA). The immunofluorescence images were acquired with ZEISS 800 confocal microscope (ZEISS, Oberkochen, Germany). The mean fluorescence intensity was measured using Image‐Pro Plus Version 6.0 software (Media Cybernetics, Silver Springs, MD, USA).

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, 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