Minute™ Yeast Mitochondria Enrichment Kit (50 Preps)

Manual & Protocol | MSDS

Traditional protocol for yeast mitochondria isolation/extraction involves a range of centrifugation-based sub-cellular 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. We feature a simple and efficient protocol for yeast mitochondria enrichment. The procedure is gentle and instrument free. Native 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.

Enrichment of Mitochondria detected by Western Blotting.  Ponceau red stained blot (A) and Western blotting (B). Lane 1, Total yeast cell lysate; Lane 2, Cytosolic fraction; Lane 3 and 4, Mitochondrial fraction from two separated Experiments. Por1 is a mitochondrial marker antibody (Thermofisher, 16G9E6BC4).

Kit includes:

References (1)

1. Yamashita, K., Miyazaki, T., Fukuda, Y., Mitsuyama, J., Saijo, T., Shimamura, S., ... & Kohno, S. (2019). The novel arylamidine T-2307 selectively disrupts yeast mitochondrial function by inhibiting respiratory chain complexes. Antimicrobial Agents and Chemotherapy, AAC-00374.