Senescence β-Galactosidase Staining Kit - Cell Signaling

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed cells for 24 hours and treat with TGF-β1 for another 24 hours.	Fix cells with 0.5 mL 1Х fixative solution for 15 min at room temperature. Add 1 ml of the β-Galactosidase Staining Solution and incubate the plate at 37°C at least overnight in a dry incubator (no CO2)

Upstream tips
Seed cells for 24 hours and treat with TGF-β1 for another 24 hours.
Protocol tips
Fix cells with 0.5 mL 1Х fixative solution for 15 min at room temperature. Add 1 ml of the β-Galactosidase Staining Solution and incubate the plate at 37°C at least overnight in a dry incubator (no CO2)

Publication protocol

In this study, cell senescence was analyzed using a Senescence-β-Galactosidase Staining kit. BmMSCs were plated in 12-well plates. Following 24 hours of culture, the cells were treated with TGF-β1 for additional 24 hours. After treatments with TGF-β1, the cells were washed twice with PBS and fixed with 0.5 mL 1Х fixative solution for 15 min at room temperature. After rinsing twice with PBS, the cells were incubated with 1 mL β-Galactosdase staining solution in a dry incubator without CO2 at 37°C overnight. The cells were imaged with a microscope when β-Galactosdase staining solution is still on the plates. The blue-dye-positive cells were viewed as senescent cells.

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