The Thermo Scientific Mitochondria Isolation Kit for Cultured Cells provides a versatile, microcentrifuge-tube method for fractionating intact mitochondria from cultured mammalian cell samples in about 40 minutes.
Features of the Mitochondria Isolation Kit for Cultured Cells:
• Fast—isolate intact mitochondria from 20 million cells in approximately 40 minutes (post-cell harvest) • Flexible—process several samples simultaneously using the reagent-based method, or obtain highest possible yield with the Dounce homogenization method • Optimizable—guidelines provided for optimizing purity vs. yield with the reagent-based method • Benchtop-easy—isolation performed in a microcentrifuge tube • Compatible—isolated mitochondria may be lysed with detergent or processed for almost any downstream application, including 2D electrophoresis
Isolation of mitochondria is typically a laborious process requiring single-sample processing with Dounce homogenization. The Mitochondria Isolation Kit uses a non-mechanical, reagent-based method that allows multiple (six) cultured cell samples to be processed concurrently in about 40 minutes. Cultured mammalian cell pellets are gently lysed using a proprietary formulation that results in maximum yield of mitochondria with minimal damage to integrity.
The product instructions describe guidelines for optimizing purity vs. yield parameters. The kit also offers an optimized Dounce homogenization procedure, which results in two-fold greater mitochondria recovery compared to the reagent-based method. Both methods use differential centrifugation to separate the mitochondrial and cytosolic fractions with a benchtop microcentrifuge and are completed in approximately 40 minutes (post-cell harvest). Once isolated, the mitochondria can be used in downstream applications such as apoptosis, signal transduction and metabolic studies, as well as to facilitate mitochondrial proteomics efforts.