Caspase-9 assay - Caspase-9 and HEK293 (Dec/13/2006 )

Hi everyone,

I would like to know if anyone has tried using a caspase-9 activity assay kit to detect apoptosis in HEK293?

I know there are other caspases which may be a better choice but the reason i chose caspase-9 because it will give me an idea of the apoptotic pathway involved, which im interested to find out later. I am inducing apoptosis with UV irradiation, which i believe that the mitochondrial pathway is involved and so caspase-9 may be appropriate (please correct me if i am wrong!)

The problem is i don't know if this assay will work in HEK293 cells and i couldn't find any relevant literature so far.

I'm a newbie in lab. Thank you for your time and attention, help is very much appreciated!

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I Hope you will find this paper very useful.

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Thx for the help, but it's not what i'm looking for.

Most papers i found usually uses HEK293 transfected with caspase-9 expressing vectors.

I'm looking for a situation whereby no caspase-9 expressing vector is involved, meaning that caspase-9 is produced in HEK293 upon apoptosis induction.

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The idea behind detecting Caspase 9 for the mitochondrial pathway is fine. And I think HEK cells should also be fine. But I am not familiar with the caspase 9 assay and its sensitivity. Induce apoptosis using chemicals or drugs from literature and try out the caspase 9 assay. Always have a positive control for apoptosis. It could be caspase 3.

Additionally, you could induce apoptosis and also try western blot to detect capase9.

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