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insect protein expression - (Mar/06/2010 )

I expressed the protein as GFP fusion protein by sf9 cell. After 96 hours infection, I saw the medium some green and then harvest the cell. then I can got green pellet. But this time I also see the medium green , but the pellet totally not green!

Is it possible that havest time is to late and the cell has already lysis and GFP protein was released into to medium?

-handsome-

handsome on Mar 6 2010, 11:24 AM said:

I expressed the protein as GFP fusion protein by sf9 cell. After 96 hours infection, I saw the medium some green and then harvest the cell. then I can got green pellet. But this time I also see the medium green , but the pellet totally not green!

Is it possible that havest time is to late and the cell has already lysis and GFP protein was released into to medium?


yes that could be possible.
do you check cell viability before you harvest your cells?

-avitas-