which ELISA-based or western blot-based apoptosis method to choose? - (Sep/30/2012 )

Hi all
I want to assess anti-proliferative effects of a plant extract on cancer cell lines. I should find a way to detect apoptosis. BUT the problem is we do not have flowcytometer here so we must ignore flow-based methods. Now I do not know which method to choose. Is one method enogh? How is caspase activity test, is there any ELISA-based method or Wester-based method available? How is the DNA ladder method? which one is more reliable? If u want to do the same thing, which methods u will select?
your urgent response is highly appreciated.
Thanks in Advance

there are many different methods. I'll list them below

- CPE detection by AO/PI. this method is very easy and it's based on morphology change. You may need a fluorescent microscope.
- DNA ladder method. DNA fragmentation kits are commercially available, but you can also collect the dead cells and extract DNA yourself. You need to run the DNA on agarose gel and if it is fragmented it means the cells are definitely dead....this is the easiest explanation I can give.
- Annexin V/PI method, which is usually by flowcytometer and you don't have it
- caspase activity. This could be done by western blot or ELISA. you need to buy antibody against caspase-3 or 9 or 8. It depends really. They are commercial kits for this method too.

I would go for DNA fragmentation first.

Curtis on Mon Oct 1 06:10:17 2012 said:

You must study the apoptotic pathways. Most people check caspase 3. thst is the last caspase in both main pathways.

Thanks again, so I will go for DNA Fragmentation 1st and then Caspase 3 activity