I had tried DNA fragmentation at a time period of 24 hrs after the treatment , but my treatment is causing early apotosis among cells that i ahve validated through various experiments, which could be the reason behind not getting DNA fragmentation as it is the marker of late apoptosis , so now i want to perform the DNA fragmentation or ladder assay but at a timeeperiod when my cells would have reached late apoptosis.
It would be really helpfull if anyone has any idea regarding this or papers where i can find such information, as through my serach of literature i could not find any information .
Thank you
My suggestion for you is to perform a time-course experiment by collecting DNA samples from your treated cells at different time points beyond 24 hr. This allows you to confirm that the treatment goes from no fragmentation to having some degree of fragmentation and then accumulates over time perhaps.
I am curious about the validation experiments that you performed which convinced you that your treatment is causing early apoptosis in the cells. What sort of experiments that you performed if you mind sharing?
Hi Trisha!
The progression from early to late apoptosis varies greatly between different cell types and/or treatment. You need to identify the time point when your cells are 'fully' dead (permeabilized) in order to properly select time point(s) for your assay.
Good luck!
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