Hi. during our study, we faced with a problem in the screening of EPG RDB 257 cell line which is a resistant cell line due to the expression of efflux pumps such as MDR 1 which actively excrete drugs out of cells. now there is a possibility that they excrete selective antibiotics (puromycin in our project) as well. what we can possibly do for this problem? I should mention our transfected vector does not have the GFP gene for monitoring the transfection results. has anyone experienced this? any guidance or solution would be appreciated.
Dear Sayedeh,
I never faced this problem and I don't know which kind of vector you transfected as I don't know this cell line. I'm just guessing, but is there a chance you could co-transfect your vector again with a plasmid containg GFP or you could check your gene/protein of interest in flow cytometry or gel electrohpesis, e.g. WB, to ensure selection of your cell line?
I just found a paper which could be of interest, as they used the same (?) cell line EPG 257/85 RDB:
Article Evaluation of transfection effectiveness using fluorescein-l...
Good luck and best regards, Irene
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