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Colorimetric Detection of Senescence-Associated β Galactosidase

Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 965)

Abstract

Most normal human cells have a finite replicative capacity and eventually undergo cellular senescence, whereby cells cease to proliferate. Cellular senescence is also induced by various stress signals, such as those generated by oncogenes, DNA damage, hyperproliferation, and an oxidative environment. Cellular senescence is well established as an intrinsic tumor suppressive mechanism. Recent progress concerning senescence research has revealed that cellular senescence occurs in vivo and that, unexpectedly, it has a very complex role in tissue repair, promoting tumor progression and aging via the secretion of various cytokines, growth factors, and enzymes. Therefore, the importance of biomarkers for cellular senescence has greatly increased. In 1995, we described the “senescence-associated β galactosidase” (SA-βgal) biomarker, which conveniently identifies individual senescent cells in vitro and in vivo. Here, we describe an updated protocol for the detection of cell senescence based on this widely used biomarker, which contributed to recent advances in senescence, aging and cancer research. We provide an example of detecting SA-βgal together with other senescence markers and a proliferation marker, EdU, in single cells.

Key words

Aging Biomarker Cellular senescence EdU labeling SA-βgal Immunostaining 
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Notes

Acknowledgments

We thank Drs. Judith Campisi and Peter de Keizer, Buck Institute for Age Research, Novato, CA, for helpful suggestions.

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Copyright information

© Springer Science+Busincess Media, LLC 2013

Authors and Affiliations

  1. 1.Duke-NUS Graduate Medical School SingaporeSingaporeSingapore
  2. 2.Department of Biochemistry and Molecular BiologyThe George Washington University Medical CenterWashingtonUSA

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