Uptake measurement in HEK cells transfected with rOCT1

1 post / 0 new
Serenity11
Serenity11's picture
Uptake measurement in HEK cells transfected with rOCT1

Hi...
Prior to doing the uptake measurements... of MPP on HEK cells transfected with rOCT1, I incubate the cells for 10minutes at 37degrees in a transfer buffer that is 1X PBS with 24.5ul MgCl2 (1M) and 45ul CaCl2 (1M). 
1. Can anyone kindly tell me why this incubation is done?
2. If suppose, I take not 1M but less conc of these solutions, say, 4mM, what can be the possible effect?
3. In the no.2 scenario is there but the incubation is done at 0 degree.. will it matter??
I will be obliged if somebody provides me with the answer... thanks..