I've been doing some MTT assays for the last couple of months, following my lab's protocol.
Then I started searching for other protocols to write a more complete one for myself and I found a difference. I would like your advice on it.
In my lab's protocol we do like this:
- culture cells and treat then as determined
- remove media of every well
- add 10 uL of 5 mg/mL MTT solution (in PBS) to each well
- incubate for 3 hours at 37°C
- remove MTT and add 100 uL of acidified isopropanol
- read at 570nm
The protocol works just fine, but in most other protocols I found that MTT solution is added TO THE MEDIA, and not TO THE CELLS as we do.
Does everyone know if this make much difference at the end result?