TUNEL - TUNEL DOESN'T WORK (May/20/2003 )

I've a problem: i analized an irradiated cell line (TK6 - 2Gy gamma rays) with TUNEL protocol but i didn't find apoptosis, but i'm sure that there're apoptosis and the positive control worked well!

I've exactky the same problem.... I worked on MEF and I'm sure they entre the apoptosis!
But no answer...

did you do the tunel by FACS or by looking under a flourescent microscope?

V

when i used to do tunel, at first i did not get through it properly. but then it is very important to standardise the time of exposure of the cells to drug or reagent before proceeding tunel. try doign it at different time intervals. that was the only thing which solved nmy problem.
all the best.