We purchased this Cell Meter™ TUNEL Apoptosis Assay Kit *Green Fluorescence* and I'm working with MDA-MB-231 cells right now trying to evaluate apoptosis in drug/polymer-drug conjugate treated cells using fluorescence microscope with TUNEL assay. The issue I'm having is that my control cells (untreated) are showing green fluorescence brighter than drug treated cells. I suppose control cells should be showing less apoptotic cells compared to anticancer drug treated one's but reverse is happening. I was wondering if someone can guide me on this issue.
Indra Putra Taufani Simply perform TUNEL assay without fixing the cells. Just incubate your drug treated cells with freshly prepared TUNEL reagent without any fixation step and proceed. Fixing can be harsh sometimes and can permeabilize the membrane resulting in exposure of nucleus to TUNEL reagent in control cells. This might help, it worked for me.
Mohammed Alvi Wow, really different protocol. Okay, I will try it.
Just for your information, I used In Situ Cell Death Detection Kit Fluorescein from Roche.
Thank you and Salam
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