SA-β-gal (Beyotime, Beijing, China) activity was determined 72 h after transfection with lentivirus. Briefly, cells were washed with PBS and fixed for 15 min. Then, cells were washed three times with PBS and stained with X-gal solution for 6–24 h at 37°C. The population of SA-β-gal-positive cells was determined by counting 100 cells per dish, and images were taken using a phase-contrast microscope at 100× magnification (Olympus, Japan). The proportions of cells positive for SA-β-gal activity are presented as a ratio of the number of positive cells to the total number of cells counted in each dish. The results are expressed as the mean of triplicates ± SD.
Sa β gal
Manufactured by Olympus
Sourced in Japan
SA-β-gal is a laboratory tool used to detect and quantify senescence-associated beta-galactosidase (SA-β-gal) activity in cells. SA-β-gal is a well-established biomarker for cellular senescence, a state of permanent cell cycle arrest. The SA-β-gal assay provides a simple and reliable method to measure this activity, which is often used in research on aging and cellular senescence.
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5 protocols using sa β gal
1
Senescence-Associated β-Galactosidase Assay
2
Senescence-Associated β-Galactosidase Assay
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HaCaT and WS1 cells were fixed in 2% formaldehyde/0.2% glutaraldehyde for 5 min at room temperature. β-Galactosidase staining solution containing X-gal (cat. no. C0602; Beyotime Institute of Biotechnology) was added after rinsing with PBS. The cells were then incubated for 6-10 h in a 37˚C incubator without CO2. Senescent cells (stained blue) were observed and images were captured using light microscopy (Olympus Corporation; magnification, x10), and positive staining areas were calculated by determining the percentage of SA-β-gal+ cells in five random fields in each of the three wells.
3
Senescence-Associated β-Galactosidase Microscopy
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For senescence associated β-galactosidase (SA-β-gal) detection microscopy analysis was performed with the inverted bright field microscope (Olympus). Fluorescence signals were detected by fluorescence microscopy (Olympus).
For statistical analysis student's t test was performed.
For statistical analysis student's t test was performed.
4
Senescence-Associated Beta-Galactosidase Assay
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For senescence associated β-galactosidase (SA-β-gal) detection microscopy analysis was performed with the inverted bright field microscope (Olympus). Fluorescence signals were detected by fluorescence microscopy (Olympus).
For statistical analysis student’s t test was performed.
For statistical analysis student’s t test was performed.
5
β-Galactosidase Senescence Assay
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HaCaT and WS1 cells were fixed in 2% formaldehyde/0.2% glutaraldehyde for 5 min at room temperature. β-Galactosidase staining solution containing X-gal (cat. no. C0602; Beyotime Institute of Biotechnology) was added after rinsing with PBS. The cells were then incubated for 6-10 h in a 37˚C incubator without CO2. Senescent cells (stained blue) were observed and images were captured using light microscopy (Olympus Corporation; magnification, x10) and positive staining areas were calculated by determining the percentage of SA-β-gal+ cells in five random fields in each of the three wells.
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