I have good results showing the induction of PARP cleavage by different agents, but I need to include a positive control for publications. Any suggestions? Thanks!
Classical inducers of apoptosis should do. We generally use cisplatin or etoposide as a positive control. Best wishes
It is depend on what stimulus your have given to your cells, for eg. if you gave cytokine treatment then FasL is suitable positive control and if your are dealing with anticancer drugs, then you have more choices like curcumin, EGCG, Cisplatin serve as positive control for PARP cleavage.
When you have already used different substances and you are looking for a simple and cheap substance I can offer you H2O2 as possible control. Reference: Activation of caspase 3 in HL-60 cells exposed to hydrogen peroxide.1999
H2O2 is a cheap and easy applicable control for early apoptosis induction for fluorescence microskopy with Hoechst 33254. This cheap stain nicely show you when the DNA starts to condense as indicator for early apoptosis.
I have used 400 or 500 microM (test from 100-700 microM with your cells) with HeLa cells and after 1 hour incubation time (37°C, 5% CO2) you see the first rounding up and chromatin condensation. After 2 hours most of the cells have condensed chromatin and after 3-4 hours most cells consists of apoptotic spheres.
You can add Hoechst directly to the culture medium to a final concentration of 5-10 µg/ml to control if it works with your cells, some are more resistant to H2O2 stess. Hoechst is cell membran premeable, just let it incubate with your cells for around 15 min (37°C, 5% CO2), then the nuclei are nicely blue and you see if it works or not.
Best Regards
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