Selectively Enrich for Mitochondrial and Cytosolic Proteomes
Overview: The Cytosol/Mitochondria Fractionation Kit (Cat. No. QIA88) is designed for extraction of cellular proteomes from mammalian cells. The simple procedure utilizes unique extraction buffers to deliver two distinct proteins fractions: a mitochondrial fraction and a cytosolic fraction. The procedure is easy to perform and does not rely upon ultracentrifugation. Protein fractions are suitable for many downstream applications including 1D and 2D gel electrophoresis, immunoblotting, and apoptotic and signal transduction-related translocation studies.
Apoptosis associated relocalization of cytochrome C from mitochondria to the cytoplasm. 107 Jurkat cells were treated (lanes 2 and 4) or untreated (lanes 1 and 3) for 7 h with 1 mM of Actinomycin D (Cat. No. 114666). Cells were harvested and processed using the Cytosol/Mitochondria Fractionation Kit (Cat. No. QIA88). 50 mg of total protein from each fraction was loaded onto a denaturing 8-12% SDS PAGE gel. The proteins were transferred to nitrocellulose and blotted with Anti-Cytochrome c Mouse mAb (Cat. No. AP1029) at 1 mg/ml. Lanes 1 and 2: Cytosolic extract. Lanes 3 and 4: Mitochondrial extract.
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