Staurosporine is a natural product derived alkaloid which is isolated from Streptomyces staurospores cultures. This protein kinase inhibitor belongs to a family of kinase inhibitors containing an indole carbozol chromophore and is a potent inhibitor of a number of kinases. The ATP binding site on these kinase enzymes appears to be targeted by these Streptomyces derived inhibitors.
Inhibition of these intracellular kinases leads to the induction of apoptosis as exhibited by the classic chromatin condensation leading to the formation of micronuclear bodies and reduced cell volumes. DNA is subsequently cleaved into oligonucleosomal fragments (laddering) as evidenced by agarose gel analysis.
Staurosporine can be used with ICT’s apoptosis and cytotoxicity detection assays to create positive controls.
1. Dissolve staurosporine powder in tissue culture grade DMSO to obtain a 1 mM staurosporine concentration.
2. 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.
3. 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.
4. 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.
5. Proceed with your experimental apoptosis induction model system.
• Molecular weight: 466.5
• Storage: -20°C