Description

Mitochondria are the powerhouse of the cells because they generate most of the supply of energy in the form of adenosine tri-phosphate (ATP). Mitochondria are double membrane organelles: an outer membrane and a folded inner membrane called cristae. Isolated mitochondria are the sample of choice to study mitochondrial respiration, assembly of the respiratory complexes, apoptosis, mtDNA and mtRNA, and mitochondrial protein profiling. The kit offers two options for the isolation of intact mitochondria. The first option is utilizing a reagent-based method allowing parallel processing of multiple samples. The second option uses traditional dounce homogenization, which provides better mitochondrial yield.

Datasheet

FAQ

Can you isolate mitochondria from frozen tissues?
We recommend using fresh tissues for mitochondria isolation- the reason is freezing procedures tend to disrupt the mitochondrial membrane and therefore it’s hard to isolate intact mitochondria from frozen tissue. However, you can isolate the mitochondria first and snap feeze them- this does not cause any decomposition of the electron transport chain components.

Can this kit be used for doing mito-activity assay in stored mitochondria, if yes then what are the storage conditions for the isolated mitochondrial?
The mitochondria isolated using K288 can be stored in the recommended buffer on the attached datasheet and stored at -80C for future use. Storage depends on which protein is going to be studied. The mitochondrial oxidoreductases are notoriously well known to have decreased activity upon prolonged storage. So the fresher the mitochondria the better.

Can the kit isolate mitochondria from blood? If so, how should the blood be collected and stored?
Yes, this kit can isolate mitochondria from PBMCs. Blood can be collected in heparinized tubes and stored at -80°C. We always prefer fresh samples to frozen and the yield and quality can differ based on the duration of storage.

Can the kit work on bacteria or yeast cells?
The kit has been standardized for mammalian cells only

What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.

Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact yopur regional BioVision distributor.

What is the shelf life of this kit?
BioVisions’s assay kits expire 6 months from the date of shipment. Some components of the kit may expire sooner as mentioned in the data sheet

Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.

Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.

Can Mitochondria prepared using K288-50 can be used for K287-100 (Cytochrome Oxidase Activity Colorimetric Assay Kit)?
Yes, mitochondria purified using K288-50 can be used with K287-100.

Can I use this kit for Mitochondria Isolation from whole blood?
K288 protocol is not optimized for whole blood. As known, RBCs do not contain mitochondria and will interfere with the assay. The option you have is to isolate PBMC or leukocytes that contain mitochondria and then use it with the kit. But again, this has not been tested here. K288 is optimized for cells and tissues.



CITATIONS

Kaowen Yan, Mitochondrial miR-762 regulates apoptosis and myocardial infarction by impairing ND2. Cell Death Dis, June 2019; 31235686.
Min Ni, Functional Assessment of Lipoyltransferase-1 Deficiency in Cells, Mice, and Humans. Cell Rep, Apr 2019; 31042466.
Maria Plataki, Mitochondrial Dysfunction in Aged Macrophages and Lung during Primary Streptococcus pneumoniae Infection is Improved with Pirfenidone. Sci Rep, Jan 2019;  30700745.
Hailong Zhao, Resveratrol induces apoptosis in human melanoma cell through negatively regulating Erk/PKM2/Bcl-2 axis. Onco Targets Ther, Dec 2018; 30588012.
Hung-Wen Liu, et al., ( 2018) Moderate Exercise Suppresses NF-κB Signaling and Activates the SIRT1-AMPK-PGC1α Axis to Attenuate Muscle Loss in Diabetic db/db Mice, Frontiers in Physiology, May. 2018, 29896118.

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