Staurosporine

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.

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.