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

oleh: Osiris Germán Idelfonso-García, Ruth Pacheco-Rivera, Brisa Rodope Alarcón-Sánchez, Jesús Serrano-Luna, Rafael Baltiérrez-Hoyos, Verónica Rocío Vásquez-Garzón, Pablo Muriel, Saúl Villa-Treviño, Julio Isael Pérez-Carreón, Jaime Arellanes-Robledo

Format: Article
Diterbitkan: Elsevier 2024-06-01

Deskripsi

Summary: 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.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.