Further, how can you prepare concentrations of bleomycin? Can somebody suggest to me how exactly I can prepare bleomycin stock solution and what is the most efficient and appropriate concentration of bleomycin that can be used to induce ds DNA damage in HEK293 cells?
Hi,
bleomicin usualy is in a liquid drug form (for example i used TEVA bleomicin 1500UI/ml). My working concentration was 0.5 mU/ml (weak exposure) and 5 mU/ml for strong exposure for HCT116 and HT29 cell culture lines. But in this way you should check if cell culture have any response to bleomicin, because due to low bleomicin permeability through cell membrane (HT29 cell line didnt respond to bleomicin at all). For dna damage response I sugest to incubate cells not more than 12 hours with the drug and check dna breaks with anti phosphoH2AX-gama ab's.
thanks Vaidotas for your valuable suggetion..
If possible can you suggest me some literature regarding my query and more specifically i am using HEK 293 cells to treat with bleomycin.
Hey Reena,
I'm looking for the same thing: concentration of bleomycin to induce DNA damage in HEK293 cells? Do you have any comments/ suggestions.
Thanks
Annie
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