Senescence β-Galactosidase Staining Kit - Cell Signaling

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	Fix cells with 2% formaldehyde/0.2% glutaraldehyde at room temperature for 10 min. Wash cells and incubate overnight at 37°C with staining solution.

Protocol tips
Fix cells with 2% formaldehyde/0.2% glutaraldehyde at room temperature for 10 min. Wash cells and incubate overnight at 37°C with staining solution.

Publication protocol

SA-β-gal activity was determined as described by Debacq-Chainiaux et al. [41]. In brief, the cells were washed twice with phosphate-buffered saline (PBS) and fixed with 2% formaldehyde/0.2% glutaraldehyde at room temperature for 10 min. After two additional washes with PBS, 1 ml of staining solution (40 mM citric acid/Na phosphate buffer, 5 mM K4[Fe(CN)6]3∙H2O, 5 mM K3[Fe(CN)6], 150 mM sodium chloride, 2 mM magnesium chloride and 1 mg/mL X-gal in distilled water, pH 6.0) were added to the cells, and incubated overnight at 37°C. The degree of cell senescence was quantified as the percentage of SA-β-gal positive cells and expressed as a percentage of the NC.

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