Protocol to detect senescence-associated β-galactosidase and immunoperoxidase activity in fresh-frozen murine tissues.

STAR Protoc

Laboratory of Liver Diseases, National Institute of Genomic Medicine - INMEGEN, Mexico City 14610, Mexico; Deputy Directorate of Humanistic and Scientific Research, National Council of Humanities, Sciences and Technologies - CONAHCYT, Mexico City 03940, Mexico. Electronic address:

Published: June 2024

Double labeling to identify different markers in the same tissue section represents a useful tool either for in situ diagnosis or characterization of molecular associations. Here, we present a protocol to detect senescence-associated β-galactosidase (SA-βGal) and immunoperoxidase (IPO) activity in fresh-frozen murine tissues. We describe steps for tissue collection, solution preparation, SA-βGal staining, IPO staining, hematoxylin counterstaining, microscopic observation, and signal quantification. This protocol can be used to detect in situ proteins alongside SA-βGal activity. For complete details on the use and execution of this protocol, please refer to Pacheco-Rivera et al..

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11017356PMC
http://dx.doi.org/10.1016/j.xpro.2024.103009DOI Listing

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