Key features and details
- Assay type: Direct
- Sample type: Suspension cells
Product nameYeast Mitochondria Isolation Kit
Sample typeSuspension cells
Species reactivityReacts with: Saccharomyces cerevisiae, Pichia pastoris
Predicted to work with: Schizosaccharomyces pombe, Candida albicans
Isolated mitochondria are useful for studying mitochondrial respiration, assembly of respiratory complexes, apoptosis, mtDNA and mtRNA, and for protein profiling.
Abcam's Yeast Mitochondria Isolation Kit (ab178779) has been tested for Pichia pastoris and Saccharomyces cerevisiae and can be used to isolate mitochondria from other yeast.
Under fermentable media, the yield is ~ 150 - 200 µg of mitochondria, and under non-fermentable media (such as glycerol), # 200 - 250 µg of mitochondria from a culture growing at OD = 20.
ab178779 is only sufficient for 25 samples.
Storage instructionsStore at -20°C. Please refer to protocols.
Components Identifier 50 tests Buffer A Blue 1 x 50ml Buffer B 1 x 50ml DTT 1 x 1ml Homogenization Buffer Amber 1 x 50ml Lysis Enzyme Mix Orange 1 x 200µl Protease Inhibitor Cocktail Red 1 vial Storage Buffer 1 x 10ml
Purified mitochondria were analyzed for intactness by using JC-1 dye, which tests the electrochemical proton gradient (ΔΨM) of the inner mitochondrial membrane. The intact purified mitochondria (control) show aggregation of JC-1 dye, whose signal can be measured at Ex/Em = 530/590 nm. Treatment with antimycin C (100 µM), an inhibitor of the electron transport chain, dissipates the mitochondrial membrane potential resulting in reduced fluorescence signal.
ab178779 has been referenced in 2 publications.
- Gao J et al. Meiotic viral attenuation through an ancestral apoptotic pathway. Proc Natl Acad Sci U S A 116:16454-16462 (2019). PubMed: 31266891
- Park SC et al. Molecular mechanism of Arabidopsis thaliana profilins as antifungal proteins. Biochim Biophys Acta Gen Subj 1862:2545-2554 (2018). PubMed: 30056100