O2k-mitochondrial preparations

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O2k-mitochondrial preparations

O2k-Procedures


O2k-Mitochondrial preparations

General remarks:

Isolation of mitochondria with well preserved function is a prerequisite for the measurement of mitochondrial respiration. In the literature a wide diversity of buffers with different composition for sample homogenization can be found as well as various settings as related to centrifugation force and time for separation of mitochondria. The appropriate method has to be selected to meet specific needs for purity and functionality of isolated mitochondria, additionally taking into account time and cost requirements of the procedure. Functional mitochondria can be prepared only from fresh and not from frozen tissue. It is pivotal that during the entire isolation procedure the samples and buffers are kept on ice.

Specific remarks:

Mitochondria from brain and liver can be separated by simple differential centrifugation of tissue homogenate. Bovine serum albumin should be used to bind free fatty acids released from membranes during the isolation procedure, as these may act as uncouplers. For isolation of mitochondria from heart and skeletal muscle, a protease should be added in the homogenization step so as to release mitochondria from the interfibrillar space.

The listed MiPNet procedures for isolation of mitochondria from rat tissues (brain, liver and heart) were evaluated in the Pharmacobiochemical Laboratory, 3rd Department of Internal Medicine, Medical Faculty, Comenius University in Bratislava, Slovak Republic as optimal for a good yield of mitochondria with satisfactory purity and well preserved function for measurements of mitochondrial respiration.


 » O2k-Catalogue: O2k-Accessories and Auxiliary Tools
ProtocolViewTopicPreparationTissue;cellOrganismLast update
MiPNet18.05 Amplex-Mouse-heartBioblast pdf »Versions
O2k-Protocols
O2k-Fluorometry: HRR and H2O2 production in mouse cardiac tissue homogenate.
HomogenateHeartMouse2019-06-24
MiPNet21.17 BloodCellsIsolationBioblast pdf  »Versions
O2k-Protocols
O2k-Protocols: Isolation of blood cells for HRR.
Permeabilized cells
Intact cells
Blood cells
Lymphocyte
Platelet
Human2018-04-09
MiPNet20.08 IsolationRatLiver-mtBioblast pdf »Versions
O2k-Protocols
Laboratory Protocol: isolation of rat liver mitochondria.
Isolated mitochondriaLiverRat2016-11-18
MiPNet20.07 IsolationRatBrain-mtBioblast pdf »Versions
O2k-Protocols
Laboratory Protocol: isolation of rat brain mitochondria.
Isolated mitochondriaNervous systemRat2016-06-18
MiPNet20.15 IsolationRatHeart-mtBioblast pdf »Versions
O2k-Protocols
Laboratory protocol: isolation of rat heart mitochondria.
Isolated mitochondriaHeartRat2016-03-25
MiPNet20.06 IsolationMouseHeart-mtBioblast pdf »Versions
O2k-Protocols
Laboratory protocol: isolation of mouse heart mitochondria.
Isolated mitochondriaHeartMouse2015-06-22
MiPNet14.14 PermeabilizedFiberPreparationBioblast pdf »Versions
O2k-Protocols contents
Preparation of permeabilized muscle fibers for diagnosis of mitochondrial respiratory function.
Permeabilized tissueHeart
Skeletal muscle
Human2015-06-22
MiPNet17.15 PBI-Shredder Mouse-heart-brain-liverBioblast pdf »Versions
O2k-Protocols
Tissue homogenates for diagnosis of mitochondrial respiratory function: Mouse heart, brain and liver.
HomogenateHeart
Nervous system
Liver
Mouse2015-06-22
MiPNet17.02 PBI-Shredder manualBioblast pdf  » Versions
O2k-Manual
PBI-Shredder HRR-Set: preparation of tissue homogenates for diagnosis of mitochondrial respiratory function.
Homogenate2015-06-19
MiPNet17.03 Shredder vs FibersBioblast pdf »Versions
O2k-Protocols
Mitochondrial respiration in permeabilized fibers versus homogenate from fish liver and heart. An application study with the PBI-Shredder.
HomogenateHeart
Liver
Fishes2015-06-18
MiPNet20.09 IsolationBeefHeart-mtBioblast pdf »Versions
O2k-Protocols
Laboratory protocol: isolation of beef heart mitochondria.
Isolated mitochondriaHeartBovines2015-06-15
MiPNet11.05 Mitos-PermeabilizedCellsBioblast pdf »Versions
O2k-Protocols
Isolated mitochondria or permeabilized tissues and cells.
Permeabilized cells
Permeabilized tissue
Isolated mitochondria
2014-04-06
MiPNet08.13 MitoIsolation-RLMBioblast pdf »Versions
O2k-Protocols
Laboratory protocol: Isolation of rat liver mitochondria.
Isolated mitochondriaLiverRat2010-08-03
PBI-Shredder O2k-Set

Auxiliary manuals

 »O2k-Catalogue: O2k-Accessories and Auxiliary Tools
ChapterViewSectionLast update
MiPNet17.02 PBI-Shredder manualBioblast pdf  » Versions
O2k-Manual
PBI-Shredder HRR-Set: preparation of tissue homogenates for diagnosis of mitochondrial respiratory function.
2015-06-19


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» O2k-Procedures: Mitochondrial and marker-enzymes