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Sample Preparation

Mitochondrial Fractionation Kit

isolate enriched mitochondrial fractions for apoptosis research

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.

 
Name Format Cat No. Price  
Mitochondrial Fractionation Kit 100 rxns 40015 $250 Buy Now

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 in 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 Mitochondrial Fractionation Kit isolates specific fractions, with little cross-contamination. (The FunctionELISA Cytochrome c Kit has since been discontinued.)

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

Contents & Storage

Each Mitochondrial Fractionation Kit contains sufficient reagents to perform 100 mitochondrial and cytosolic extractions from a 100 mm tissue culture dish of cells grown to confluence.