Senescence Cells Histochemical Staining Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed 1×10^5 and treat with C-1305	Fix with with 2% formaldehyde and 0.2% glutaraldehyde in PBS for 5 min. Add 1 ml of the Staining Mixture per well and incubate at 37 °C without CO2 until the cells are stained blue

Upstream tips
Seed 1×10^5 and treat with C-1305
Protocol tips
Fix with with 2% formaldehyde and 0.2% glutaraldehyde in PBS for 5 min. Add 1 ml of the Staining Mixture per well and incubate at 37 °C without CO2 until the cells are stained blue

Publication protocol

Senescence-associated (SA) beta-galactosidase histochemical staining in the CHO cells was performed after exposure to C-1305 as described by Dimri et al15. Following C-1305-treatment, the cells (1×105) were washed twice with PBS and fixed with 2% formaldehyde and 0.2% glutaraldehyde in PBS for 5 min. The cells were then washed again with PBS and stained with a solution of 1 mg/mL 5-bromo-4-chloro-3-indolyl-β-D-galactosidase in dimethylformamide (20 mg/mL stock), 5 mmol/L potassium ferrocyanide, 5 mmol/L potassium ferricyanide, 150 mmol/L NaCl, 40 mmol/L citric acid/sodium phosphate, pH 6.0, and 2 mmol/L MgCl2. Following overnight incubation at 37 °C, the cells were washed twice with PBS and representative fields were photographed under a light microscope (Olympus BX60, 20×objective).

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