Senescence Detection Kit - Biovision

Protocol tips

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	Fix the cells with 0.5 ml of Fixative Solution for 10 - 15 min at room temperature Add 0.5 ml of the Staining Solution Mix to each well. Cover the plate. Incubate overnight at 37°C.

Protocol tips
Fix the cells with 0.5 ml of Fixative Solution for 10 - 15 min at room temperature Add 0.5 ml of the Staining Solution Mix to each well. Cover the plate. Incubate overnight at 37°C.

Publication protocol

RNA interference-mediated silencing of TRPM8 induced replicative senescence. The BxPC-3 and PANC-1 cells were transfected with either anti-TRPM8 siRNA or non-targeting control siRNA and then seeded at 4 x 104 cells / 2 ml medium in each well of a 12-well cell culture cluster (Corning Inc., costar). The transfected cells were incubated at 37°C for 72 h, and then analyzed for activity of SA β-gal using the Senescence Detection Kit (Biovision Medical Products) according to the manufacturer’s instructions. Images were acquired under an inverted light microscope with phase contrast (Nikon TE-300) and processed using Adobe Photoshop CS3 extended. Appearance of blue color indicates SA β-gal activity (arrows), which is detected in the anti-TRPM8 siRNA-transfected cells but not in those transfected with control siRNA.

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Manufacturer protocol

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