Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

Native BrdU staining

XW Xiao Wu
BW Bin Wang
request Request a Protocol
ask Ask a question
Favorite

Cells were labeled with BrdU (10 μM) for 36 h53, washed with culture medium after labeling, and treated with CPT (1 μM, 1 h). Cells were then pre-extracted with pre-extraction buffer, fixed with 3% paraformaldehyde/2% sucrose solution, treated with cold methanol for 20 min at −20 °C, washed with PBS, and treated with cold acetone for 30 s. After blocking with 2% BSA for 1 h, staining was performed with BrdU antibody followed by Alexa-488-conjugated secondary antibody. Images were captured with 80i eclipse Nikon microscope using ×40 objective. BrdU nuclear intensity was measured by ImageJ software (ImageJ 2.0 and 64 bit Java8 from https://imagej.nih.gov).

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.

0/150

tip Tips for asking effective questions

+ Description

Write a detailed description. Include all information that will help others answer your question including experimental processes, conditions, and relevant images.

post Post a Question
0 Q&A