Mitochondrial (Tissue) Isolation Kit
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|Product Overview :||With Mitochondrial (Tissue) Isolation Kit, it is possible to isolate coupled mitochondria* from freshly harvested mammalian heart, liver, and kidney tissue as long as precautionary steps are taken. The presence of trace amounts of detergent can have detrimental effects on mitochondrial membrane integrity. Therefore, when using Mitochondrial (Tissue) Isolation Kit it is important to ensure that all glassware (and polycarbonate centrifuge tubes), that will come into contact with isolation buffers, have been rinsed free of detergents. To remove detergents from glassware, wash using 70% ethanol in place of soap, followed by several rinses with deionized water. Glassware should then be rinsed in UltraPure water. It is strongly recommended that plasticware be avoided. All steps of this preparation are carried out on ice or at 4°C. When handling glassware containing tissue homogenates or crudely isolated mitochondria, take care not to wrap hands around the glassware. Heat from hands can be easily transferred to mitochondria, potentially compromising membrane integrity. Should membrane integrity be compromised due to the presence of detergents or heat, mitochondrial protein can still be isolated; however the quality of the resulting prep may be poor (i.e., uncoupled, poor respiration rate).
*Coupled mitochondria are defined as mitochondria capable of phosphorylating ADP at the F1FO-ATP synthase (Complex V) through the utilization of a proton (H+) gradient, which is generated by the electron transport chain (ETC) in the presence of appropriate substrates (e.g., succinate) and in the absence of uncouplers and mitochondrial inhibitors.