Abstract
Identifying molecules that serve as markers for cell aging is a goal that has been pursued by several groups. Senescence-associated β galactosidase (SA-βgal) staining is broadly used and very easily detected. β-gal is a lysosomal enzyme strongly correlated to the progression of cell senescence. Here, we describe a simple, fast, and quantitative protocol to quantify SA-βgal activity in cell lysate extracts by a chemiluminescent method using galacton as substrate.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Lee BY, Han JA, Im JS, Morrone A, Johung K, Goodwin EC, Kleijer WJ, DiMaio D, Hwang ES (2006) Senescence-associated beta-galactosidase is lysosomal beta-galactosidase. Aging Cell 5:187–195
Campisi J (2011) Cellular senescence: putting the paradoxes in perspective. Curr Opin Genet Dev 21:107–112
Bronstein I, Martin CS, Fortin JJ, Olesen CE, Voyta JC (1996) Chemiluminescence: sensitive detection technology for reporter gene assays. Clin Chem 42:1542–1546
Jain VK, Magrath IT (1991) A chemiluminescent assay for quantitation of beta-galactosidase in the femtogram range: application to quantitation of beta-galactosidase in lacZ-transfected cells. Anal Biochem 199:119–124
Dimri GP, Lee X, Basile G, Acosta M, Scott G, Roskelley C, Medrano EE, Linskens M, Rubelj I, Pereira-Smith O et al (1995) A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc Natl Acad Sci U S A 92:9363–9367
Shlush LI, Itzkovitz S, Cohen A, Rutenberg A, Berkovitz R, Yehezkel S, Shahar H, Selig S, Skorecki K (2011) Quantitative digital in situ senescence-associated beta-galactosidase assay. BMC Cell Biol 12:16
Kurz DJ, Decary S, Hong Y, Erusalimsky JD (2000) Senescence-associated (beta)-galactosidase reflects an increase in lysosomal mass during replicative ageing of human endothelial cells. J Cell Sci 113(Pt 20):3613–3622
Noppe G, Dekker P, de Koning-Treurniet C, Blom J, van Heemst D, Dirks RW, Tanke HJ, Westendorp RG, Maier AB (2009) Rapid flow cytometric method for measuring senescence associated beta-galactosidase activity in human fibroblasts. Cytometry A 75:910–916
Yang NC, Hu ML (2004) A fluorimetric method using fluorescein di-beta-d-galactopyranoside for quantifying the senescence-associated beta-galactosidase activity in human foreskin fibroblast Hs68 cells. Anal Biochem 325:337–343
Debacq-Chainiaux F, Erusalimsky JD, Campisi J, Toussaint O (2009) Protocols to detect senescence-associated beta-galactosidase (SA-betagal) activity, a biomarker of senescent cells in culture and in vivo. Nat Protoc 4:1798–1806
Gary RK, Kindell SM (2005) Quantitative assay of senescence-associated beta-galactosidase activity in mammalian cell extracts. Anal Biochem 343:329–334
Bassaneze V, Miyakawa AA, Krieger JE (2008) A quantitative chemiluminescent method for studying replicative and stress-induced premature senescence in cell cultures. Anal Biochem 372:198–203
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Busincess Media, LLC
About this protocol
Cite this protocol
Bassaneze, V., Miyakawa, A.A., Krieger, J.E. (2013). Chemiluminescent Detection of Senescence-Associated β Galactosidase. In: Galluzzi, L., Vitale, I., Kepp, O., Kroemer, G. (eds) Cell Senescence. Methods in Molecular Biology, vol 965. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-239-1_9
Download citation
DOI: https://doi.org/10.1007/978-1-62703-239-1_9
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-238-4
Online ISBN: 978-1-62703-239-1
eBook Packages: Springer Protocols