Hello Philip Drell

There are other methods such as

1. You may try an antibiotic selection method wherein you introduce your gene of interest into HEK293 cells along with a selectable marker gene (e.g., neomycin resistance or puromycin resistance) on the same plasmid or a separate plasmid using a co-transfection strategy. After transfection, incubate the cells in the medium containing the appropriate antibiotic. Only cells that have successfully integrated the selectable marker gene into their genome will survive the antibiotic selection process.

2. Another way would be to co-transfect HEK293 cells with your gene of interest and a plasmid expressing a fluorescent protein (e.g., GFP, mCherry). After transfection and a period of incubation, you may use flow cytometer to sort cells based on fluorescence. Cells expressing the fluorescent protein are sorted and collected.

3. You may also try the functional assay wherein if your gene of interest encodes a protein with a detectable function (e.g., enzymatic activity), you can use a functional assay to select for cells expressing the protein of interest. After transfection, perform the functional assay on the cells. Cells that show the expected activity are selected.

Best,

Philip Drell

A simple, professional way to select HEK293 cells after transfection—no microscope needed at the hood:

Notes

• Perform all media changes inside a biosafety cabinet using sterile technique; you don’t need a microscope there.
• If your transfection was transient and the plasmid lacks a resistance gene, antibiotic selection won’t work—re-transfect with a vector that includes the selectable marker or use a co-transfection strategy with a resistance plasmid.

Just a fyi if you're transfecting cells with your protein of interest + fluorescent protein, probably will be a good idea to image by microscopy, to ensure your protein is localizing correctly. It could also be a good idea to expand out multiple clones such as those with high, medium, low expression of your fluorescent fusion protein (when viewing by microscopy). Again in cases where you are putting in a fluorescent FP