Apoptosis Assay

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Grainne
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Apoptosis Assay

I am trying to design an experiment to look at apoptosis in adherent cells whihc have been transfected with my protein of interest. I am inducing apoptosis using cycloheximide, but i am unsure of how much to use, and do i induce all cells, or just use the cycloheximide to induce some cells as a control. I have 18 wells seeded, 6 which have been transfected, half of these will also be stimulated to induce an inflammatory response also. I would appreciate any help

Satinder
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Grainne
Grainne
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Thank You, i cant acess that

Thank You, i cant acess that paper at the minute, subscribtion has run out. How long do i induce with the cycloheximide,

heehawmcduff
heehawmcduff's picture
I've also seen a lot of

I've also seen a lot of people use combination treatments of cycloheximide and Fas-ligand to induce apoptosis.  To give you some idea, the concentration of Fas ligand used was 1ug/ml and the concentration of cycloheximide was 2ug/ml and the cells were incubated for 12 hours before measurement of apoptosis.

For cycloheximide alone, I would use a slightly higher conc over a couple of different time points i.e. 6 hours and 12 hours.

Grainne
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Thank you, i will try that.

Thank you, i will try that.

Satinder
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send me yr email id. I will

send me yr email id. I will sens the article through mail.

Jason King
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Satinder,

Satinder,
If you have a protocol that is not already in the "Protocols"  area, it would be great if you could add it.

Thanks

PM