Mitochondria are the power house 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 is a useful tool to study mitochondrial respiration, assembly of the respiratory complexes, apoptosis, mtDNA and mtRNA, and for protein profiling. BioVision’s Yeast Mitochondria Isolation kit will enable fast and easy purification of mitochondria from yeast cells, utilizing yeast cell wall lysis and homogenization. This Mitochondria Isolation Kit is tested for Pichia pastoris & Saccharomyces cerevisiae & can be used to isolate mitochondria from other yeast. Under fermentable media, the yield is ~150-200 µg of mitochondria & under non-fermentable media (e.g. Glycerol) ~200-250 µg of mitochondria from a culture of OD ~20.
|#Cat + Size||K259-50 (Size: 50 assays)|
|Applications||Isolation of highly pure intact mitochondria from yeast cells. Mitochondrial respiration studies, assembly of the complexes, apoptosis, mtDNA and mtRNA, and for protein profiling. Western blot and ELISA|
|Features & Benefits||• Simple & convenient protocol
• The kit provides unique formulations of ready-to-use buffers and reagents to isolate intact mitochondria
|Kit Components||• Buffer A • Buffer B • 1 M DTT • Homogenization Buffer • Lysis Enzyme Mix • Storage Buffer • Protease Inhibitor Cocktail|
|Shipping Conditions||Gel Pack|
|USAGE||For Research Use Only! Not For Use in Humans.|
Please let me know tools alternative for "glass douncer"
The Barocycler, the PCT Shredder has been used by others(http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3172370/). But we recommend the dounce or other glass/Teflon homogenizer.
In isolation process, does mitochondria content leakage happen?
No, we have tested the mitochondrial integrity and we haven’t see any leakage
Is it possible to quantify molecules (nucleotide, ATP, amino acid and so on) in isolated yeast mitochondria? Yes you can. Here are some of our suggested applications: • Mitochondrial respiration studies, assembly of the complexes, apoptosis, mtDNA and mtRNA, and for protein profiling • Western blot and ELISA
Can the kit work with yeasts other than Saccharomyces cerevisiae and Pichia pastoris?
Theoretically, the kit should work with yeasts having similar cell wall structures.
Could you tell me the pH of the storage buffer and if there is sugar included to keep isotonic?
The storage buffer has a neutral pH and does contain sugar.
I can see that the yield of mitochondria is ~150-200ug under fermentable media. What is the yield per procedure?
We typically use OD600 to measure the cell number of yeast cells in culture. Typically an OD600=1 is approximately equal to 3*10⁷cells/ml. 200-250ug of mitochondria can be isolated from approximately 6-10*10⁸cells/ml.
I was wondering if mitochondria isolated from this product can be used in mitochondrial DNA immunoprecipitation assay (ChIP in mitochondrial DNA). Mitochondria isolated using this kit can be used for many downstream applications including ChIP. However for ChIP applications we recommend using freshly isolated mitochondria.
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.
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Our products such as Elisa, Antibodies, Proteins, Peptides and sequencing kits are covered by Biolinkk quality warranty and will work as described in datasheet, a free replacement or money back is guaranteed if does not perform according to datasheet.