Minute™ Mitochondria Isolation Kit for Mammalian Cells and Tissues (50 Preps)

Manual & Protocol | Material Safety Data Sheets (MSDS)

Minute™ Mitochondria Isolation Kit for Mammalian Cells and Tissues comprises optimized buffers and protein extraction filter cartridges with 2.0 ml collection tubes. The kit is designed to rapidly isolate native mitochondria proteins from cultured mammalian cells or tissues. Due to protein extraction filter cartridges, the protein isolation procedure is simple and provides a high yield. The process can be completed in less than 30 minutes. Unlike many commercial mitochondria preparation kits, this kit offers a wide range of starting cells and isolated mitochondria. The buffers are detergent and EDTA-free. A Dounce homogenizer or a tissue blender is not needed.

Figure 1. Enrichment of Mitochondrial Marker (Ubiquinol-Cytochrome C Reductase Core Protein) with Minute™ Mitochondria Isolation Kit. 

 A. SDS-PAGE profiles of total protein extract vs. isolated mitochondrial proteins. Lanes 1, 3, 5, total proteins from isolated mouse kidney cells, mouse skeletal muscle and liver tissues, respectively. Lanes 2, 4, 6, isolated mitochondrial proteins from mouse kidney cells, mouse skeletal muscle and liver tissues, respectively.

 B. Western blotting. Proteins in A were transferred to nitrocellulose membranes and probed with anti-ubiquinol-cytochrome C reductase core protein (ab96333, abcam, Cambridge, MA) and anti-lamin B1 (ab16048, a nuclear envelope marker protein. abcam, Cambridge, MA). 

 C. Densitometry measurement of mitochondrial marker signals.

Kit includes:

1. Gou, R. et al. (2014). Resveratrol suppresses oxidized low-density lipoprotein-induced macrophage apoptosis through inhibition of intracellular reactive oxygen species generation, lox-1 and the p38 MAPK pathway. Cellular Physiology and Biochemistry. 34:603-616.
2. Hu, H., Zhu, W., Qin, J., Chen, M., Gong, L., Li, L., ... & Ye, D. (2016). Acetylation of PGK1 Promotes Liver Cancer Cell Proliferation and Tumorigenesis. Hepatology.
3. Gao, G., Wang, Z., Lu, L., Duan, C., Wang, X., & Yang, H. (2017). Morphological analysis of mitochondria for evaluating the toxicity of α-synuclein in transgenic mice and isolated preparations by atomic force microscopy. Biomedicine & Pharmacotherapy.
4. Zhang, L., Liu, J., Zhou, F., Wang, W., & Chen, N. (2018). PGC-1α ameliorates kidney fibrosis in mice with diabetic kidney disease through an antioxidative mechanism. Molecular medicine reports.
5. Wu, Y., Gao, W. N., Xue, Y. N., Zhang, L. C., Zhang, J. J., Lu, S. Y., ... & Sun, L. K. (2018). SIRT3 aggravates metformin-induced energy stress and apoptosis in ovarian cancer cells. Experimental Cell Research.
6. Shen, L., Sun, B., Sheng, J., Yu, S., Li, Y., Xu, H., ... & Sun, L. (2018). PGC1α promotes cisplatin resistance in human ovarian carcinoma cells through upregulation of mitochondrial biogenesis. International Journal of Oncology.
7. Han, W., Cao, F., Gao, X. J., Wang, H. B., Chen, F., Cai, S. J., ... & Ding, H. Z. ZIC1 acts a tumor suppressor in breast cancer by targeting survivin. International Journal of Oncology.
8. Ouyang, J., Zeng, Z., Fang, H., Li, F., Zhang, X., & Tan, W. (2019). SIRT3 inactivation promotes acute kidney injury through elevated acetylation of SOD2 and p53. Journal of Surgical Research, 233, 221-230.
9. Zhou, K., Yao, Y. L., He, Z. C., Chen, C., Zhang, X. N., Yang, K. D., ... & Niu, Q. (2018). VDAC2 interacts with PFKP to regulate glucose metabolism and phenotypic reprogramming of glioma stem cells. Cell Death & Disease, 9(10), 988.
10. Xue, Y. N., Yu, B. B., Li, J. L., Guo, R., Zhang, L. C., Sun, L. K., ... & Li, Y. (2018). Zinc and p53 Disrupt Mitochondrial Binding of HK2 by Phosphorylating VDAC1. Experimental Cell Research.
11. Chen, X. Y., Ren, H. H., Wang, D., Chen, Y., Qu, C. J., Pan, Z. H., ... & Li, D. F. (2019). Isoliquiritigenin Induces Mitochondrial Dysfunction and Apoptosis by Inhibiting mitoNEET in a Reactive Oxygen Species-Dependent Manner in A375 Human Melanoma Cells. Oxidative Medicine and Cellular Longevity, 2019.
12. Ma, J., Chen, L., He, X. X., Wang, Y. J., Yu, H. L., He, Z. X., ... & Zhu, X. J. (2019). Functional prediction and characterization of Dip2 gene in mice. Cell biology international.
13. Cai, M., He, P., & Fang, D. L. Hypoxia‑induced mitochondrial translocation of DNM1L increases mitochondrial fission and triggers mPTP opening in HCC cells via activation of HK2. Oncology Reports.
14. Guo, Z., Song, T., Xue, Z., Liu, P., Zhang, M., Zhang, X., & Zhang, Z. (2019). Using CETSA assay and a mathematical model to reveal dual Bcl-2/Mcl-1 inhibition and on-target mechanism for ABT-199 and S1. European Journal of Pharmaceutical Sciences, 105105.
15. Yue, J., Shen, Y., Liang, L., Cong, L., Guan, X., Li, Z., ... & Xu, W. In situ and ex situ surface‐enhanced Raman spectroscopy (SERS) analysis of cell mitochondria. Journal of Raman Spectroscopy.
16. Duan, C., Kuang, L., Xiang, X., Zhang, J., Zhu, Y., Wu, Y., ... & Li, T. (2020). Activated Drp1-mediated mitochondrial ROS influence the gut microbiome and intestinal barrier after hemorrhagic shock. Aging (Albany NY), 12(2), 1397.
17. Duan, C., Kuang, L., Xiang, X., Zhang, J., Zhu, Y., Wu, Y., ... & Li, T. (2020). Drp1 regulates mitochondrial dysfunction and dysregulated metabolism in ischemic injury via Clec16a-, BAX-, and GSH-pathways. Cell Death & Disease, 11(4), 1-19.
18. Han, W., Zhang, C., Shi, C. T., Gao, X. J., Zhou, M. H., Shao, Q. X., ... & Yuan, J. L. (2020). Roles of eIF3m in the tumorigenesis of triple negative breast cancer. Cancer Cell International, 20(1), 1-16.
19. Fei, Q., Ma, H., Zou, J., Wang, W., Zhu, L., Deng, H., ... & Wang, N. (2020). Metformin protects against ischaemic myocardial injury by alleviating autophagy-ROS-NLRP3-mediated inflammatory response in macrophages. Journal of Molecular and Cellular Cardiology.
20. Hu, X., Xia, M., Wang, J., Yu, H., Chai, J., Zhang, Z., ... & Sun, L. (2020). Dual PI3K/mTOR inhibitor PKI-402 suppresses the growth of ovarian cancer cells by degradation of Mcl-1 through autophagy. Biomedicine & Pharmacotherapy, 129, 110397.
21. Lu, D., Yang, Z., Xia, Q., Gao, S., Sun, S., Luo, X., ... & Li, X. ACADSB regulates ferroptosis and affects the migration, invasion and proliferation of colorectal cancer cells. Cell Biology International.
22. Deng, X., Lin, N., Fu, J., Xu, L., Luo, H., Jin, Y., ... & Su, J. (2020). The Nrf2/PGC1α Pathway Regulates Antioxidant and Proteasomal Activity to Alter Cisplatin Sensitivity in Ovarian Cancer. Oxidative Medicine and Cellular Longevity, 2020.
23. Wang, Y., Kang, Y., Qi, C., Zhang, T., Zhao, H., Ji, X., ... & Shi, G. (2020). Pentoxifylline enhances antioxidative capability and promotes mitochondrial biogenesis for improving age-related behavioral deficits. Aging, 12.
24. Huang, J., Xie, P., Dong, Y., & An, W. (2020). Inhibition of Drp1 SUMOylation by ALR protects the liver from ischemia-reperfusion injury. Cell Death & Differentiation, 1-19.
25. Luo, Q., Wu, X., Zhao, P., Nan, Y., Chang, W., Zhu, X., ... & Liu, Z. (2021). OTUD1 Activates Caspase‐Independent and Caspase‐Dependent Apoptosis by Promoting AIF Nuclear Translocation and MCL1 Degradation. Advanced Science, 2002874.
26. She, H., Zhu, Y., Deng, H., Kuang, L., Fang, H., Zhang, Z., ... & Li, T. (2021). Protective effects of dexmedetomidine on the vascular endothelial barrier function by inhibiting mitochondrial fission via ER/mitochondria contact. Frontiers in Cell and Developmental Biology, 9.
27. Zhou, J., Zhao, Y., Guo, Y. J., Zhao, Y. S., Liu, H., Ren, J., ... & Ji, E. S. (2021). A rapid juvenile murine model of nonalcoholic steatohepatitis (NASH): Chronic intermittent hypoxia exacerbates Western diet-induced NASH. Life Sciences, 119403.
28. Yu, S., Yan, X., Tian, R., Xu, L., Zhao, Y., Sun, L., & Su, J. (2021). An Experimentally Induced Mutation in the UBA Domain of p62 Changes the Sensitivity of Cisplatin by Up-Regulating HK2 Localisation on the Mitochondria and Increasing Mitophagy in A2780 Ovarian Cancer Cells. International Journal of Molecular Sciences, 22(8), 3983.
29. Wu, X., Li, J., Wang, S., Jiang, L., Sun, X., Liu, X., ... & Yang, G. (2021). 2-Undecanone Protects against Fine Particle-Induced Kidney Inflammation via Inducing Mitophagy. Journal of Agricultural and Food Chemistry.
30. Zhao, L., Liu, D., Ma, W., Gu, H., Wei, X., Luo, W., & Yuan, Z. (2021). Bhlhe40/Sirt1 Axis-Regulated Mitophagy Is Implicated in All-Trans Retinoic Acid-Induced Spina Bifida Aperta. Frontiers in Cell and Developmental Biology, 9, 1025.
31. Feng, S. T., Wang, Z. Z., Yuan, Y. H., Wang, X. L., Guo, Z. Y., Hu, J. H., ... & Zhang, Y. (2021). Inhibition of dynamin-related protein 1 ameliorates the mitochondrial ultrastructure via PINK1 and Parkin in the mice model of Parkinson’s disease. European Journal of Pharmacology, 174262.
32. 
    Wang, Y., Zhang, T., Zhao, H., Qi, C., Ji, X., Yan, H., ... & Shi, G. (2021). Pentoxifylline Enhances Antioxidative Capability and Promotes Mitochondrial Biogenesis in D-Galactose-Induced Aging Mice by Increasing Nrf2 and PGC-1α through the cAMP-CREB Pathway. Oxidative Medicine and Cellular Longevity, 2021.
    
33. Duan, C., Kuang, L., Hong, C., Xiang, X., Liu, J., Li, Q., ... & Li, T. (2021). Mitochondrial Drp1 recognizes and induces excessive mPTP opening after hypoxia through BAX-PiC and LRRK2-HK2. Cell Death & Disease, 12(11), 1-10.
34. Li, T., Zhao, G., Zhang, T., Zhang, Z., Chen, X., Song, J., ... & Weng, C. (2021). African Swine Fever Virus pE199L Induces Mitochondrial-Dependent Apoptosis. Viruses, 13(11), 2240.
35. Meng, H., Sun, L. K., Su, J., Yan, W. Y., Jin, Y., Luo, X., ... & Wang, H. L. (2021). Serine protease HtrA2/Omi regulates adaptive mitochondrial reprogramming in the brain cortex after ischemia/reperfusion injury via UCP2-SIRT3-PGC1 axis. Human Cell, 1-20.
36. Li, X., Hou, P., Ma, W., Wang, X., Wang, H., Yu, Z., ... & He, H. (2021). SARS-CoV-2 ORF10 suppresses the antiviral innate immune response by degrading MAVS through mitophagy. Cellular & Molecular Immunology, 1-12.

37. Yan, X., Qu, X., Liu, B., Zhao, Y., Xu, L., Yu, S., ... & SU, J. (2021). Autophagy-induced HDAC6 activity during hypoxia regulates mitochondrial energy metabolism through the β-catenin/COUP-TFII axis in hepatocellular carcinoma cells. Frontiers in Oncology, 4275. 

38. Xiao, L., Hu, Q., Peng, Y., Zheng, K., Zhang, T., Yang, L., ... & Liu, Y. (2021). TRAP1 suppresses oral squamous cell carcinoma progression by reducing oxidative phosphorylation metabolism of Cancer-associated fibroblasts. BMC cancer, 21(1), 1-14.
39. Cheng, M., Yu, H., Kong, Q., Wang, B., Shen, L., Dong, D., & Sun, L. (2022). The Mitochondrial PHB2/OMA1/DELE1 Pathway Cooperates with Endoplasmic Reticulum Stress to Facilitate the Response to Chemotherapeutics in Ovarian Cancer. International Journal of Molecular Sciences, 23(3), 1320.

40. Liu, J., Li, L., Xie, P., Zhao, X., Shi, D., Zhang, Y., ... & Li, T. (2022). Sevoflurane induced neurotoxicity in neonatal mice links to a GSK3β/Drp1-dependent mitochondrial fission and apoptosis. Free Radical Biology and Medicine.

41. Huang, F., Tang, X., Ye, B., Wu, S., & Ding, K. (2022). PSL-LCCL: a resource for subcellular protein localization in liver cancer cell line SK_HEP1. Database, 2022.

42. He, X. T., Li, X., Zhang, M., Tian, B. M., Sun, L. J., Bi, C. S., ... & Chen, F. M. (2022). Role of molybdenum in material immunomodulation and periodontal wound healing: Targeting immunometabolism and mitochondrial function for macrophage modulation. Biomaterials, 121439.
43. Zhao, M., Wei, H., Li, C., Zhan, R., Liu, C., Gao, J., ... & Zheng, L. (2022). Gut microbiota production of trimethyl-5-aminovaleric acid reduces fatty acid oxidation and accelerates cardiac hypertrophy. Nature Communications, 13(1), 1-18.
44. Hu, W., Guo, Y., Wang, X., Cui, T., Li, C., Liu, R., & Yin, C. (2022). Angiotensin-(1–7) promotes mitochondrial translocation of human telomerase reverse transcriptase in HUVECs through the TOM20 complex. Archives of Biochemistry and Biophysics, 109218.
45. Xu, Y., Chen, R., Yan, J., Zang, G., Shao, C., & Wang, Z. (2022). CD137 Signal Mediates Cardiac Ischemia–Reperfusion Injury by Regulating the Necrosis of Cardiomyocytes. Journal of Cardiovascular Translational Research, 1-13.
46. Li, B., & Liu, L. (2022). Fibroblast growth factor 21, a stress regulator, inhibits Drp1 activation to alleviate skeletal muscle ischemia/reperfusion injury. Laboratory Investigation, 1-10.
47. Wang, X. L., Feng, S. T., Wang, Y. T., Zhang, N. N., Guo, Z. Y., Yan, X., ... & Zhang, Y. (2022). Mangiferin, a natural glucoxilxanthone, inhibits mitochondrial dynamin-related protein 1 and relieves aberrant mitophagic proteins in mice model of Parkinson's disease. Phytomedicine, 154281.
48. Wu, T., Jia, L., Lei, S., Jiang, H., Liu, J., Li, N., ... & Lei, L. (2022). Host HSPD1 Translocation from Mitochondria to the Cytoplasm Induced by Streptococcus suis Serovar 2 Enolase Mediates Apoptosis and Loss of Blood–Brain Barrier Integrity. Cells, 11(13), 2071.
49. An, W., Zheng, M., Ai, Z., & Chen, Y. (2022). Imbalance in ALR ubiquitination accelerates the progression of nonalcoholic steatohepatitis to hepatocellular carcinoma.
50. Wang, M., Zhang, B., Zhang, C., Zhang, X., Tang, S., Sun, G., & Sun, X. (2022). Quantitative Crotonylome Analysis Reveals the Mechanism of Shenkang Injection on Diabetic Nephropathy. Oxidative Medicine and Cellular Longevity, 2022.
51. Wang, Y., Hu, L. F., Liu, N. H., Han, M. M., Zhou, T. J., Xing, L., & Jiang, H. L. (2022). Suppression of mitochondrial heterogeneity via engineered mitochondria for reversion of mitochondrial disease-related phenotypes.
52. Zhu, Y., Lei, L., Wang, X., Chen, L., Li, W., Li, J., ... & Li, X. (2023). The E3 ubiquitin ligase NEDD4-1 protects against acetaminophen-induced liver injury by targeting VDAC1 for degradation. Acta Pharmaceutica Sinica B.
53. Edwards-Hicks, J., Apostolova, P., Buescher, J. M., Maib, H., Stanczak, M. A., Corrado, M., ... & Pearce, E. L. (2023). Phosphoinositide acyl chain saturation drives CD8+ effector T cell signaling and function. Nature Immunology, 1-15.
54. Yin, M., Dong, J., Sun, C., Liu, X., Liu, Z., Liu, L., ... & Deng, H. (2023). Raddeanin A Enhances Mitochondrial DNA‐cGAS/STING Axis‐Mediated Antitumor Immunity by Targeting Transactive Responsive DNA‐Binding Protein 43. Advanced Science, 2206737.