- Dissolve staurosporine powder in tissue culture grade DMSO to obtain a 1 mM staurosporine concentration.
- Prepare 50 – 100 µL aliquots of the DMSO solubilized staurosporine stock solution and store them frozen at < -20°C. A frozen vial of staurosporine may only be rethawed 2X before it must be discarded.
- Spike cell cultures at a staurosporine concentration of 1 µM in the cell culture media. This equates to a 1 µL spike of the 1 mM staurosporine stock per mL of the cell culture suspension. This concentration works well for inducing many cell lines including HL-60 and Jurkat cells when using a 4-5 hour 37°C incubation period. Typical cell suspension concentrations that have been used for this staurosporine induction protocol range from 1 x 105 – 1 x 106 cells/mL.
- Perform time course studies on your particular cell line to ascertain the optimal staurosporine concentration and exposure time required to achieve good apoptosis induction levels in your experimental system.
- Proceed with your experimental apoptosis induction model system.
Question: I already purchased your FAM-FLICA kit for flow staining. I am trying to use Staurosporine to induce apoptosis as a positive control. Since the cells I am using are not cell line in culture, they are actually flushed out of mouse lungs. How should I use Staurosporine to induce apoptosis? Any method and advice to help?
Answer: Staurosporine is a potent protein kinase inhibitor and is very effective at inducing apoptosis across a range of cell lines and types. I understand you are not working with a particular cell line, but instead are culturing cells after they have been flushed out of mouse lungs. I would suggest setting up an experiment where you evaluate a range of staurosporine concentrations and exposure periods. As a starting point, we normally use 1 µM concentration for 4 hours to induce apoptosis in >90% of Jurkat cells. We have found that some cell lines, such as U-937 cells, may require up to 6 hours for similar results. Thus it is best to set up a titration experiment with your cells and derive the optimal staurosporine concentration and exposure time experimentally.