293 Cells: how to transfect?

[Dr Andrew J. Doherty]

HEK293 cells are notorius for not sticking to anything! We use them to 
transfect with Glutamate receptor subunits and stick them to glass 
coverslips coated with poly-D-lysine. They seem to be OK then. They can 
be pathched, used in confocal microscopy and immunocytochemistry (live) - 
so they will stick! But they are very sensitive to their environment. 
They don't like being at room temperature when growing on ordinary tissue 
culture plasticware and can die very easily if they don't like your 
transfection methodology. We use a CaPO4 method, with overnight 
transfection incubations, and they seem fine with this.

Hope this is helpful

Andy Doherty
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