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Live stain with propidium iodide - Incubation time & temperature? (Mar/22/2010 )

I'm going to do a live stain using propidium iodide. Any recommendations regarding incubation time and temperature? I was going to incubate at 4C for 30 min using 10 M PI in PBS. However, I've heard that after some time, PI will diffuse even through intact cell membranes, so might 30 min be too long? Are 5-10 min long enough?

-Aquifex-

After reading some not so helpful papers and doing some trial and error on my own, I've come to the conclusion that adding the PI directly before measurement works just fine. No incubation required, in fact incubating the cells for more than 10-15 minutes is probably already too long. I used a final concentration of 10 g / mL in ice-cold PBS with about 2,000,000 cells / mL.

-Aquifex-

Aquifex on Mar 25 2010, 08:18 AM said:

After reading some not so helpful papers and doing some trial and error on my own, I've come to the conclusion that adding the PI directly before measurement works just fine. No incubation required, in fact incubating the cells for more than 10-15 minutes is probably already too long. I used a final concentration of 10 g / mL in ice-cold PBS with about 2,000,000 cells / mL.


Hi, I currently add 5ul of 1mg/ml propodium iodide (Invitrogen) to my 100ul cell suspension (about 200,000 cells), keep in Rt for 15mins before read. It works fine with me.

-goldfinger-

please can you tell me for how long can i keep propiudium iodide sol. that i've prepared?

-pana-