Mitochondrial Isolation: A Technique to Isolate Mitochondria from Human Ovarian Cancer Tissue Sample

Published: April 30, 2023

Abstract

Source: Zhan, X. et al. Preparation of Mitochondria from Ovarian Cancer Tissues and Control Ovarian Tissues for Quantitative Proteomics Analysis. J. Vis. Exp. (2019)

This video describes the procedure for isolating mitochondria from human ovarian cancer tissues using differential-speed centrifugation in combination with density gradient centrifugation. The isolated mitochondria can be used for proteomic analysis of human ovarian cancer mitochondrial proteome.

Protocol

All procedures involving human participants have been performed in compliance with the institutional, national, and international guidelines for human welfare and have been reviewed by the local institutional review board. 1. Preparation of mitochondria from human ovarian cancer tissues Ovarian tissue samples including ovarian cancer tissues (n = 7) were used for this protocol. Prepare 250 mL of the mitochondrial isolation buffer by mixi…

開示

The authors have nothing to disclose.

Materials

Diamine tetraacetic acid (EDTA) Sigma 798681-100G Anhydrous, free-flowing, Redi-Dri, ≥98%
DTT Sigma 10197777001 1,4-Dithiothreito
Ethylen glycol bis(2-aminoethyl ether)tetraacetic acid (EGTA) Sigma E0396-10G BioXtra, ≥97 .0%
Homogenizer SilentShake HYQ-3110
Low-temperature super-speed centrifuger Eppendorf 5424R
Centrifuge XiangYi TDZ4–WS
Mannitol Macklin M813424-100G Mannitol is a polyol (polyhydric alcohol) produced from hydrogenation from fructose that functions as a sweetener, humectant, and bulking agent. It has low hygroscopicity and poor oil solvency.
Phenylmethanesulfonyl fluoride (PMSF) protease inhibitor Solarbio P0100-1ML PMSF is a protease inhibitor that reacts with serine residues to inhibit trypsin, chymotrypsin, thrombin, and papain.
Potassium chloride Macklin P816354-25G Potassium chloride, KCI, also known as potassium muriate and sylvite, is a colorless crystalline solid with a salty taste that melts at 776°C (1420 OF). It is soluble in water, but insoluble in alcohol. Potassium chloride is used in fertilizers, pharmaceuticals, photography, and as a salt substitute

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記事を引用
Mitochondrial Isolation: A Technique to Isolate Mitochondria from Human Ovarian Cancer Tissue Sample. J. Vis. Exp. (Pending Publication), e20370, doi: (2023).

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