Target preparation plating cell

Each of these assays has drawbacks associated with them. The major obstacle for developing rapid screens for inhibitors of cytochrome c release from mitochondria is that enriched mitochondria have a finite time in which they can be used. We are in the process of testing mitochondria preparations to determine the stability of the mitochondria with respect to use in the cytochrome c release assay. Mitochondria stored on ice for 4 h (the longest time we have tested at the time of this writing) can be used in the cytochrome c release assay. Therefore, it is possible to run multiple assay cycles with one preparation of mitochondria. The short format cytochrome c release assay is not affected by cell permeability, makes no assumptions about the mechanism of action, does not use cultured cells, is targeted at the process to be inhibited (i.e., cytochrome c release from mitochondria), and is a colormetric assay easily monitored by spectrophotometric plate readers. This assay would, therefore, increase the chances of detecting lead compounds to be subsequently modified to increase potency, cell permeability, and pharmacological efficacy. 

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