Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

SA-β-gal staining

HJ Hee-Young Jeon
SH Seok Won Ham
JK Jun-Kyum Kim
XJ Xiong Jin
SL Seon Yong Lee
YS Yong Jae Shin
CC Chang-Yong Choi
JS Jason K. Sa
SK Se Hoon Kim
TC Taehoon Chun
XJ Xun Jin
DN Do-Hyun Nam
HK Hyunggee Kim
request Request a Protocol
ask Ask a question
Favorite

To detect cell senescence, non-irradiated or irradiated GBM cells and xenograft tumor sections (4 μm thick) were stained using a Senescence β-Galactosidase Staining Kit (9860, Cell Signaling Technology) according to manufacturer instructions. Briefly, after washing the plate or slides with PBS, a 1× fixative solution was added to each well or slide for 15 min at room temperature. Next, the plates or slides were rinsed with PBS, and then β-galactosidase staining solution was added. The plates or slides were incubated overnight at 37 °C in a dry incubator (no CO2). After incubation, the slides were stained with Nuclear fast red (NFR; N3020, Sigma-Aldrich) for 5 min and rinsed with tap water. Cells and tissues were analyzed using an inverted fluorescence microscope (Axio Observer D1, Carl Zeiss) to detect the blue staining.

Copyright and License information: The Author(s) ©2019
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