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Cayman’s MitoCheck® Mitochondria (Cell Culture) Isolation Kit enables the isolation of functionally intact mitochondria from cultured cells in less than 60 minutes. Cells are initially resuspended in an isotonic buffer and subjected to mechanical disruption to liberate organelles. This is followed by sequential differential centrifugation to first remove nuclei and undisrupted cells and subsequently pellet mitochondria.
An optional digitonin cell permeabilization reagent is also provided to facilitate plasma membrane disruption, which may enable increased mitochondrial yields. Digitonin mediates selective disruption of cholesterol-rich membranes, resulting in plasma membrane permeabilization, while mitochondrial membranes, which are comparatively low in cholesterol, remain intact.1
This kit provides sufficient reagents to perform up to 60 isolations. Mitochondria isolated using this kit are functionally intact and may be used immediately for respirometry studies or other downstream application. Additional Cayman MitoCheck® assay kits may be used to characterize mitochondrial function.
WARNING This product is not for human or veterinary use.
1. Permeabilized cell and skinned fiber techniques in studies of mitochondrial function in vivo. Mol. Cell Biochem. 184(1-2), 81-100 (1998).