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The Mitochondrial Fractionation Kit is designed to facilitate the study of apoptotic and signal transduction pathways. It provides a unique formulation of buffers and reagents that enable isolation of highly enriched mitochondrial and cytosolic fractions from various mammalian cell lines, including both apoptotic and non-apoptotic cells. The fractionation procedure is simple and easy to perform. No ultracentrifugations are required and no toxic chemicals are involved.
The Mitochondrial Fractionation Kit's high-quality reagents and optimized protocol eliminate cross-contamination and produce high yields of properly segregated mitochondrial & cytosolic fractions (Figure 1). Extracts are ideal for use with the Active Motif’s FunctionELISA™ Cytochrome c Kit, or in other procedures that require mitochondrial extract, like gelshift assays, Western blot analysis, ELISA or other assays.
The Mitochondrial Fractionation Kit advantage
- Quality-controlled reagents ensure reproducibility
- No need to optimize your own procedure
- Complete kit contains all required reagents
- Prepare both the cytosolic & mitochondrial fractions
Figure 1: Localization of cytochrome c in HeLa cells.
HeLa cells were grown to 90-95% confluence and treated with 10 µM Actinomycin D. Cells were harvested at 0 hours and after 16 hours of induction. Mitochondrial and cytosolic extracts were isolated using the Mitochondrial Fractionation Kit. Two µg of each lysate was tested using the FunctionELISA™ Cytochrome c Kit (A) and Western blot analysis using a 4-20% Tris-Glycine gel (B). The results demonstrate that the kit isolates specific fractions, with little cross-contamination.
Lane 1: Cytosolic extract, no induction
Lane 2: Mitochondrial extract, no induction
Lane 3: Cytosolic extract, 16 hr post-induction
Lane 4: Mitochondrial extract, 16 hr post-induction
Lane 5: Protein standard
