Senescence Cells Histochemical Staining Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed cells fro 72 hours and washe twice with PBS, Fix for 6 min with fixation buffer containing 2% formaldehyde, 0.2% glutaraldehyde, 7.0 mM Na2HPO4, 1.5 mM KH2PO4, 140 mM NaCl, and 2.68 mM KCl	Add 1 ml of the Staining Mixture per well and incubate overnight at 37 °C without CO2 until the cells are stained blue.

Upstream tips
Seed cells fro 72 hours and washe twice with PBS, Fix for 6 min with fixation buffer containing 2% formaldehyde, 0.2% glutaraldehyde, 7.0 mM Na2HPO4, 1.5 mM KH2PO4, 140 mM NaCl, and 2.68 mM KCl
Protocol tips
Add 1 ml of the Staining Mixture per well and incubate overnight at 37 °C without CO2 until the cells are stained blue.

Publication protocol

In order to test for senescence, AsPC-1 and BxPC-3 cells treated with TTFields for 72 h, were washed twice with PBS, fixed for 6 min with fixation buffer containing 2% formaldehyde, 0.2% glutaraldehyde, 7.0 mM Na2HPO4, 1.5 mM KH2PO4, 140 mM NaCl, and 2.68 mM KCl and stained with senescence cells histochemical staining kit (CS0030, SigmaeAldrich) according to manufacturer instructions. After an overnight incubation at 37 C, images of the cells were taken using Nikon Eclipse T100 light microscope equipped with Nikon DC-Fi1c camera (Nikon instruments, Tokyo Japan). Analysis of the images was carried by a researcher blinded to the treatment group.

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