BestProtocols: Pharmacological Induction of Apoptosis with Camptothecin
Note: This protocol is intended for use with the specific products mentioned within it. Substituting different products is not recommended.
- A cell line or primary cells susceptible to apoptosis induction.
- RPMI-1640 medium supplemented with 10% FCS
- 1 mM stock solution of Camptothecin prepared in DMSO
- Tissue culture flasks or tissue culture plates
- Prepare cells in fresh RPMI-1640 medium with 10% FCS at a concentration of 0.5 x 106 cells/mL in desired tissue culture flasks or tissue culture plates.
- Add an appropriate amount of 1 mM Camptothecin to the cell suspension to achieve a final concentration of 4-6 µM. The negative control should consist of cells maintained in medium with an equivalent dilution of DMSO only.
- Incubate cells for the amount of time optimal for your cell type in a humidified, 5% CO2 incubator at 37°C. It is recommended that you first do a time course to get an idea of how sensitive you cells are to undergo apoptosis
- Harvest cells by centrifugation and proceed with appropriate assay to evaluate the induction of apoptosis.
- Note: Other pharmacological reagents that have been shown to induce apoptosis include: Actinomycin D, Aphidocolin, Cycloheximide, Dexamethasone, 5-Fluorouracil, Hydroxyurea, and Staurosporine.
Traganos F, Seiter K, Feldman E, Halicka HD, Darzynkiewicz Z. 1996. Induction of apoptosis by camptothecin and topotecan. Ann N Y Acad Sci. 13:803:101-10.
Morris EJ, Geller HM. 1996. Induction of neuronal apoptosis by camptothecin, an inhibitor of DNA topoisomerase-I: evidence for cell cycle-independent toxicity. J Cell Biol. 1996 Aug;134(3):757-70.
For Research Use Only. Not for use in diagnostic procedures.