Protocol tips
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Protocol tips |
Downstream tips |
|
Fix cells in 2% paraformaldehyde and 0.02% glutaraldehyde in PBS
Add 1 ml of the Staining Mixture per well and incubate for 16 h at 37 °C without CO2 until the cells are
stained blue |
|
Protocol tips |
Fix cells in 2% paraformaldehyde and 0.02% glutaraldehyde in PBS
Add 1 ml of the Staining Mixture per well and incubate for 16 h at 37 °C without CO2 until the cells are
stained blue |
Publication protocol
SA‐β‐gal‐positive cells were detected applying the Senescence Cells Histochemical Staining Kit obtained from Sigma‐Aldrich. In brief, cells were fixed with 2% paraformaldehyde and 0.02% glutaraldehyde in PBS and incubated for 16 hr in staining solution containing X‐Gal (5 mM C6N6FeK4/5 mMC6N6FeK3/2 mM MgCl2/1 mg/ml 5‐bromo‐4‐chloro‐3‐indolyl‐β‐d‐galacto‐pyranoside in PBS). SA‐β‐gal positive cells as percentage of total cell counts were calculated using an Olympus IX50 inverted microscope (Olympus, Center Valley, PA). For photometrical SA‐β‐gal detection, cells were fixed and incubated in staining solution supplemented with 0.2 mM fluorescein di‐β‐d‐galactopyranoside (FDG) 24 hr before fluorescence determination (excitation wave length: 485 nm, emission wave length: 535 nm).
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Manufacturer protocol
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