Qproteome Mitochondria Isolation Kit
For efficient isolation of high-purity mitochondria preparations
- High-purity, intact mitochondria
- Highly reproducible using a standardized procedure
- Convenient, fast, and easy-to-use kit format
Cat. No. / ID: 37612
The Qproteome Mitochondria Isolation Kit provides fully active mitochondrial preparations of very high purity in a fast, reproducible, and standardized procedure that uses a standard bench-top centrifuge.
One of the major problems facing researchers working with mitochondria is efficiently separating mitochondrial and cytosolic fractions. Traditional preparation methods are tedious, time-consuming, and require access to an ultracentrifuge. The Qproteome Mitochondria Isolation Kit provides fully active mitochondrial preparations of very high purity in a fast, reproducible, and standardized procedure that uses a standard bench-top centrifuge.
Mitochondria are the subject of intense study in a number of fields, including apoptosis, proteomics, biomedical research, and drug discovery. The Qproteome Mitochondria Isolation Kit delivers high-purity mitochondria whose proteins retain their native conformation and full biological activity — making them highly suited for apoptosis studies, 2D-PAGE analysis, and other biochemical assays.
Mitochondria were isolated using the Qproteome Mitochondria Isolation Kit or another commercially available kit (Supplier S) according to manufacturer’s instructions. Mitochondrial membrane integrity is indicated by formation of fluorescent aggregates following uptake of a carbocyanine dye.
|applications||SDS-PAGE, mass spectrometry, activity assays|
|samplesize||5 x 10e6 - 2 x 10e7 cells, 20 mg - 60 mg|
The Qproteome Cell Compartment Kit, the Qproteome Nuclear Protein Kit, the Qproteome Mitochondria Isolation Kit, and the PhosphoProtein Purification Kit are compatible with tissue samples. The respective protocol can be found in the respective handbook.
The Qproteome Mammalian Protein Prep Kit is also compatible with tissue, there is the QIAGEN Supplementary Protocol: Purification of protein from animal tissues using the Qproteome Mammalian Protein Prep Kit and the TissueRuptor™ available. For all these protocols the TissueRuptor is used.
We have not tested this application. We would suggest to split the sample, and perform different fractionation procedures separately on the two sample portions.
If splitting the sample is not feasible, then it should theoretically be possible to use the pellet generated at step 7 of the QProteome Mitochondria Isolation Kit protocol (containing cell debris, nuclei and unbroken cells) as starting material for the Qproteome Nuclear Protein Kit.
Please note that this application will have to be tested and optimized by the enduser.
Highly pure and intact mitochondria can be collected after the density gradient centrifugation at step 11b of the Qproteome Mitochondria Isolation protocol.
The fraction collected at step 9 of the protocol, prior to the cushion centrifugation, contains intact as well as broken mitochondria, and may also comprise structures of the ER, Golgi and plasma membranes.
The Qproteome Mitochondria Isolation Kit can be used with 5 x 106 to 2 x 107 cells. Depending on the cell line, the yield from a single fractionation procedure is 20–80 µg protein from 5 x 106 cells.
Protein can be isolated from the mitochondria using, e.g. the QProteome Mammalian Protein Prep Kit, or home-made lysis buffers.
The Qproteome Mitochondria Isolation Kit has been developed for cultured mammalian cells. We have tested HeLa, Cos7, NIH, HEK 293 and SH-SY5Y cell lines.
We have not tested animal tissue, yeast, or plant cells. Animal tissue might work if a method for cell disruption that leaves the mitochondria intact can be found. Unfortunately, we do not have any protocol recommendations for these starting materials.