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SAOS.2 Cell detachment without using Trypsin. - (Nov/01/2013 )

Hi, I am trying to detach cells from a 24-well plate for analysis by atomic absorption where I will be looking for Calcium in the samples. For this reason I must ensure that I minimize the amount of external Calcium added to my samples. I have been trying to detach the cells using concentration HCl however this does not seem to be working. Any other suggestions?

 

-yp41-

Do you need the cells intact, or is it OK if they are lysed?  If lysed, how about just scraping them into some Ca2+ free PBS?

-bob1-

What cell line are you using? I work with HEK293T cells and simple up/down pipetting unsticks the cells for me from the plastic. 

-gvbdxz-

293 are notorious for being weakly attached, this is not the case for the majority of cell lines, where simply pipetting will not work at all.

-bob1-