The kit is intended for the isolation of intact mitochondria, so most of the isolated mitochondria will contain intact inner and outer membranes. While trace amounts of mitochondria-associated membranes (MAMs) may co-purify during the process, the protocol is not optimized to enrich or preserve MAMs. Please see the Product Information Sheet at the link below for further details.
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/204/819/mitoiso1pis-ms.pdf
MITOISO1
Mitochondria Isolation Kit
sufficient for 10-20 g (animal tissue), sufficient for 50 assays (2 mL), isolation of enriched mitochondrial fraction from animal tissues
Sinonimo/i:
Mitochondria Purification Kit
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About This Item
Prezzi e disponibilità al momento non sono disponibili
Prodotti consigliati
Livello qualitativo
impiego
sufficient for 10-20 g (animal tissue)
sufficient for 50 assays (2 mL)
Durata
2 yr
tecniche
fractionation: suitable
Condizioni di spedizione
dry ice
Temperatura di conservazione
−20°C
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Applicazioni
Mitochondria Isolation Kit has been used:
- for the separation of mitochondrial and cytosolic fraction for determining the branched-chain aminotransferase (BCAT) activity[1]
- for the isolation of mitochondria from glomeruli[2] and heart[3]
- for the isolation of mitochondria from injured brain samples for the measurement of mitochondrial transmembrane potential[4]
The Mitochondria Isolation Kit provides a fast and easy isolation of an enriched mitochondrial fraction from animal tissues as well as the testing of the electrochemical proton gradient (ΔΨ) of the inner mitochondrial membrane. Useful for mitochondria mediated apoptosis studies.
Caratteristiche e vantaggi
- Specially formulated extraction reagents & proven procedure suitable for small scale isolation - Isolate an enriched, intact mitochodrial fraction in a microfuge tube.
- Produces functionally active, intact mitochondria - Resulting mitochondria are suitable for in vitro assays for apoptosis, oxidative stress or other studies.
- Includes stain & protocol for determining inner membrane integrity - Determine the integrity of the inner mitochondrial membrane without the need to purchase other reagents.
- Compatible with the Cytochrome c Oxidase Assay Kit - Allows easy determination of the integrity of the outer mitochondrial membrane
I componenti del kit sono disponibili anche separatamente
N° Catalogo
Descrizione
SDS
- T9201Trypsin from bovine pancreas, powder, ≥7,500 BAEE units/mg solidSDS
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Indicazioni di pericolo
Consigli di prudenza
Classi di pericolo
Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
Organi bersaglio
Respiratory system
Codice della classe di stoccaggio
10 - Combustible liquids
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Articoli
Centrifugation separates organelles based on size, shape, and density, facilitating subcellular fractionation across various samples.
Cellular apoptosis assays to detect programmed cell death using Annexin V, Caspase and TUNEL DNA fragmentation assays.
Cellular oxidative stress is countered by enzymatic scavengers and antioxidant modulators against reactive oxygen species damage.
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Is it possible to use the MITOISO1 kit to also isolate MAMs?
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Where is the step to get cytosol?
1 answer-
The kit is intended for the isolation of intact mitochondria. It was not developed for cytoplasma collection. Nevertheless, reviewing the protocol, it can be assumed that the cytoplasmic fraction should be in the supernatant after the second centrifugation (point 8 of the Preparation of mitochondria from soft tissues protocol). Please keep in mind that this is not tested for suitability for cytoplasmic fraction.
Please see product MIT1000 Mitochondria/Cytosol Fractionation Kit for efficient isolation of intact mitochondria from cultured cells. This kit can be used for tissue samples as long as the tissue is first homogenized into a single cell suspension. It may be helpful to use the same protocol that is recommended for cells following homogenization of the tissue into a single cell suspension in the supplied Isotonic Mitochondrial Buffer containing protease inhibitors, then follow the kits protocol for cells: https://www.sigmaaldrich.com/product/mm/mit1000.
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Are the extraction buffers from Product MITOISO1, Mitochondria Isolation Kit, available separately?
1 answer-
The extraction buffers supplied with this kit are only available with the kit.
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When using Product MITOISO2, Mitochondria Isolation Kit, how can I store my isolated mitochondria?
1 answer-
Please see below for information on how best to store your isolated mitochondria using the MITOISO2 kit:If only interested in the proteins isolated from mitochondria, storage at -20 deg C is recommended. If intact mitochondria are required, storage at 4 deg C is recommended. These intact mitochondria are stable after refrigeration for up to 24 hours.
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When using Product MITOISO1, Mitochondria Isolation Kit, how long can I store my isolated mitochondria?
1 answer-
If only interested in the proteins isolated from mitochondria, storage at -20 deg C is recommended. If intact mitochondria are required, storage at 4 deg C is recommended. These intact mitochondria are stable after refrigeration for up to 24 hours.
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What is the Department of Transportation shipping information for this product?
1 answer-
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
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How much tissue can I use with Product MITOISO1, Mitochondria Isolation Kit,?
1 answer-
The procedures described in the bulletin are for small amounts of tissue samples (50-200 mg). Larger amounts of tissue can be purified with modifications listed in the technical bulletin. We recommend a minimum of 50 mg.
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Is the Product MITOISO1, Mitochondria Isolation Kit, JC-1 stain available separately?
1 answer-
Yes, while the component J4519 is not available separately, JC-1 stain is listed separately as Sigma Product No. T4069 or Product No. 40882.
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Can Product MITOISO1, Mitochondria Isolation Kit, be used for isolation of mitochondria from cultured cells?
1 answer-
It is possible to use this kit for cultured cells, however MITOISO2 is a kit that was specifically designed for this purpose. This kit was originally designed for isolation from tissues.
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