Senescence Detection Kit - Merck

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed 1.5×10^5 cells/well	Pretreat with 4 µM PAB for 3 days. 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.

Upstream tips
Seed 1.5×10^5 cells/well
Protocol tips
Pretreat with 4 µM PAB for 3 days. 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

The MCF-7 cells (1.5×105 cells/well) were seeded into 24-well culture plates (Nalge Nunc International, Penfield, NY, USA). Following incubation overnight, 4 µM PAB was added and the plates were incubated for 3 days. The senescence detection kit was used according to the manufacturer's instructions, as previously described (6). Finally, the cells were observed under a phase contrast microscope (Olympus IX51; Olympus Corporation, Tokyo, Japan) for the development of a blue color.

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Manufacturer protocol

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