In order to treat a cell culture with TSA for re-expression of some genes without direct cytotoxic effect.
Hello there; the concentration needed will very much depend on the cell type so I would try doing a concentration response curve in an in vitro cell-based assay, followed up by a cell viability assay. Generally though, TSA is very potent and will frequently, if not usually, kill cells in the uM ranges. You'll probably find that something in the 10-100 nM-range does the job, if not possibly even less. Once you've got your highest in vitro dose that doesn't kill the cells, check the effect on gene expression by doing an acetyl histone H3/H4 western blot (the acetyl-histone antibodies from Millipore/Upstate are generally very good). Hope that might help, good luck!
Hello there; the concentration needed will very much depend on the cell type so I would try doing a concentration response curve in an in vitro cell-based assay, followed up by a cell viability assay. Generally though, TSA is very potent and will frequently, if not usually, kill cells in the uM ranges. You'll probably find that something in the 10-100 nM-range does the job, if not possibly even less. Once you've got your highest in vitro dose that doesn't kill the cells, check the effect on gene expression by doing an acetyl histone H3/H4 western blot (the acetyl-histone antibodies from Millipore/Upstate are generally very good). Hope that might help, good luck!
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