Senescence β-Galactosidase Staining Kit - Cell Signaling

Reporter gene assay β-galactosidase substrates - HEK293 human embryonic kidney cells

Experiment
Reporter gene assay β-galactosidase substrates - HEK293 human embryonic kidney cells
Product
Senescence β-Galactosidase Staining Kit - Cell Signaling from Cell Signaling Technology
Manufacturer
Cell Signaling Technology

Protocol tips

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)
Downstream tips
Visualise cells after 11 to 14 hr,

Publication protocol

The senescent cells were detected with senescence β-galactosidase staining kit (Cell Signaling, Beverly, MA, USA) according to manufacturer’s protocol. The growth media was removed and the cells were once rinsed with PBS. Next, a fixative solution containing 2% formaldehyde and 0.2% glutaraldehyde was added to the cells. Then, again the cells were rinsed with PBS two times and incubated with a staining solution containing 5-bromo-4-chloro-3-indolyl-D-galactopyranoside, citric acid/sodium phosphate (pH 6.0), 1.5 M NaCl, 20 mM MgCl2, potassium ferrocyanide and potassium ferricyanide. After 11 to 14 hr, light microscopy was applied to identify the senescent cells which can be visualized as blue cells. Thirty fields were randomly selected to calculate the percentage of SA-β-gal cells.



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Manufacturer protocol

Download the product protocol from Cell Signaling Technology for Senescence β-Galactosidase Staining Kit - Cell Signaling below.

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