Tripsin EDTA detachment - doubt (Aug/01/2005 )
I was trying to detach the monolayer by Trypsin EDTA. I could not pellet it out after centrifugation . it is still in the for of a clumped mass in the falcon tube. I used 2000 rpm for 15 mnts. Anyone got some comment on this.
try simply not to spin the cells. serum in the medium will "kill" all the trypsin so there is no need to remove it.
but in general i'm trying not to spin the cells to fast - usually ~1000 rpm, 5 min.
you do not need to remove the trypsin as long as you use serum containing medium. Adding medium at least twice the volume of the trypsin-EDTA solution will be enaugh to desactivate it.
Hope it helps
i never saw that. For spinning cells, recommendations are no more than 250-300g. So check the rpm of your centrifuge.
But i agree with the replies. Medium is supposed to inactivate trypsin. For that, i inactivate trypsin by use at least 3volume of medium for1vol of trypsin.
Thanks a lot everybody