Hello all,
I am performing telomerase inhibition experiments that will result in replicative senescence in cancer cell culture. Can anyone recommend assays to use to measure replicative senescence in cell culture? Thanks.
Measuring the SA-B-gal activity, incorporation of BrdU (IF staining), WB for p21 or/and p16 induction, p-p53 and p53 expression, staining for 53BP1 and gamma H2AX (IF; DNA damage). And last but not least - enlarged flattened morphology (eg - staining the actin with phalloidin).
You can measure the Beta-galactosidase activity, there is a simple colorimetric kit from Thermo: Thermo Scientific Mammalian β-Galactosidase Assay Kit. Also western blots for cell cycle proteins such as p21 and p53 to measure cell cycle arrest which is a hallmark of senescent cells. You should also see that the proliferative capacity of your cell is markedly reduced.
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