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Traditional protocols for yeast mitochondria isolation/extraction involve a range of centrifugation-based

subcellular fractionation procedures. Typically the techniques include spheroplast preparation, glass-bead

lysis using a homogenization instrument, differential centrifugation and several density gradient procedures

using a variety of gradient media with ultracentrifugation. The procedures are very tedious and time

consuming. Here we feature a simple and rapid protocol for yeast mitochondriaenrichment. The procedure is

gentle and instrument-free. Nativ mitochondrial proteins can be isolated from yeast in about one hour without

ultracentrifugation. This kit contains optimized detergent-free protein extraction buffers. The protein yield is in

the range of 150-250 μg/sample. The materials provided are sufficient for 50 extractions.