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Conveniently enrich the mitochondrial or cytosolic subcellular fractions from mammalian cells or tissues using the Mitochondrial Isolation Kit. This kit offers a quick mitochondrial isolation technique that has been verified for detection using western blot analysis and may be used to study apoptosis and signaling events between the two fractions.
Features:
• Fast and efficient enrichment of the active mitochondrial subcellular fraction
• Verified for use with western blot analysis
The Mitochondrial Isolation Kit offers two options to isolate the mitochondrial subcellular fraction. The first option is a reagent-based method for processing up to six samples concurrently. The second method utilizes Dounce homogenization, providing a two-fold increase in isolated mitochondria from a single sample. Both methods employ microcentrifugation to separate the mitochondrial fraction from the cytosolic fraction.
The enriched mitochondrial fractions preserve their biological activity and are compatible with downstream applications, including the study of mitochondrial respiration, apoptosis, inner mitochondrial membrane potential, and protein profiling (Itahana & Zhang Cancer Cell, 2008).
Figure 1. Western Blot of Mitochondrial Markers After Mitochondrial Isolation from Cultured Cells
Mitochondria were isolated from HEK293 cells with Mitochondrial Isolation Kit using Dounce homogenization. Mitochondrial (M) and cytosolic (C) fractions were analyzed via western blot for two mitochondrial proteins: voltage-dependent anion channels (VDAC) and cytochrome C. Results demonstrate that mitochondria remain intact after enrichment.
Figure 2. Mitochondria Isolated Using the Mitochondrial Isolation Kit Maintain Healthy Mitochondrial Function
Mitochondria were isolated from HEK293 cells using Mitochondrial Isolation Kit and stained with JC-1 (Iodide) dye (Catalog #100-0993) in the presence (grey bar) or absence (orange bar) of valinomycin. High levels of fluorescence indicate that healthy mitochondrial function is retained after isolation.
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