Senescence β-Galactosidase Staining Kit - Cell Signaling

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed 3 × 10 cells per well.	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 3 × 10 cells per well.
Protocol tips
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

Beta-galactosidase staining was performed using a senescence-associated β-galactosidase (SA-β-Gal) staining kit (Cell Signaling Technology, Danvers, MA) for 12 hours. Briefly, human adipose-derived stem cells cultured at passages 5, 10, 15, and 20 with or without REAC exposure were cultured in 6-well plates (3 × 10 per well) for 12 hours, fixed with fixative solution, and then processed according to the kit instructions. All the experiments were repeated three times, and one of the representative set of results is shown. The cells were then photographed under an inverted microscope at 100 × magnification for qualitative detection of SA-β-Gal activity. The numbers of positive (blue) and negative cells were counted in five random fields under the microscope (at 200 × magnification and bright field illumination), and the percentage of SA-β-Gal-positive cells was calculated as the number of positive cells divided by the total number of cells counted.

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