Hi Guys, hope all is well with you. I want to overexpress a protein using HEK 293 Cell line. Could you please kindly tell me what is the most convenient way to do so. I read something about finding a clone for the protein gene, could you kindly clarify on how to do so. Many thanks for your time.
Hi Moe,
You start with a plasmid that contains cDNA of the protein you want to over express. Some of cDNA are quite cheap but some can be more than 2k dollars. Sometimes you will have to clone out your cDNA of interest.
Once you have plasmid with the clone, you need to transfect cells. Hek293 are easy to transfect, I use lipofectamin or prime jet reagent. After this you need to do antibiotic selection in order to generate a new cell line that will over express the protein of interest and characterize it. Whole process takes aprox.. 2 months.
hope this helps. If you need details about any particular step I might be able to provide you with a protocol.
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