Mitochondrial Extraction Kit (NBP2-29448)

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Western Blot: Mitochondrial Extraction Kit [NBP2-29448] - Mouse liver cells were fractionated using . 20 ug of cytosolic (lane 1) and mitochondrial (lane 2) extracts were probed with anti-Cytochrome C antibody, and ...read more

Product Details

Summary
Reactivity MuSpecies Glossary
Kit Type
Extraction Kit

Order Details

Mitochondrial Extraction Kit Summary

Kit Type
Extraction Kit

Applications/Dilutions

Application Notes
The procedure described here for the isolation of mitochondria is based on the principle of differential centrifugation. The high density nuclei are first removed by low-speed centrifugation on a sucrose cushion. The supernatant containing the mitochondria is subjected to high speed centrifugation to retrieve the mitochondria. This kit contains reagents required for the isolation of mitochondria from 10 grams of mammalian tissues (liver, muscle, etc.) or 100 reactions for cells grown in tissue culture plates. The procedure describes isolation of mitochondria from 1 gram of starting tissue material (mouse liver) or 5 x 107 HeLa cells. The Mitochondrial and Cytosolic fractions can be used for studying apoptotic and signal transduction pathways to detect translocation of any factors of interest within the two fractions by western blotting, ELISA, or other assay.
Publications
Read Publications using
NBP2-29448 in the following applications:

  • WB
    9 publications

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID: 27392939)

Packaging, Storage & Formulations

Storage
Storage is content dependent.

Kit Components

Components
  1. 1M DTT 100 ul
  2. 100X Protease Inhibitor Cocktail (PIC) 100 ul
  3. 10X PBS 2 x 50 ml
  4. 5X Suspension Buffer 2 x 50 ml
  5. 5X Homogenization Buffer 50 ml
  6. Mitochondria Lysis Buffer 50 ml
  7. Mitochondrial Staining Solution 1 ml

Notes

Addition Equipment Required
1 Sterile Water
2 Standard balance (tissues)
3 High-speed centrifuge and compatible centrifuge tubes
4 Teflon homogenizer
5 Cell scraper
6 Microcentrifuge tubes

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Kits are guaranteed for 6 months from date of receipt.
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Publications for Mitochondrial Kit (NBP2-29448)(15)

We have publications tested in 2 confirmed species: Human, Mouse.

We have publications tested in 1 application: WB.


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WB
(9)
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Human
(1)
Mouse
(5)
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Showing Publications 1 - 10 of 15. Show All 15 Publications.
Publications using NBP2-29448 Applications Species
Chakraborty S, Ghosh S, Banerjee B et al. Mephebrindole, a synthetic indole analog coordinates the crosstalk between p38MAPK and eIF2a/ATF4/CHOP signalling pathways for induction of apoptosis in human breast carcinoma cells Apoptosis 2016 Jul 08 [PMID: 27392939] (Mouse) Mouse
Tadi SK, Sebastian R, Dahal S et al. Microhomology Mediated End Joining is the Principal Mediator of Double-strand Break Repair During Mitochondrial DNA Lesions. Mol. Biol. Cell. 2015 Nov 25 [PMID: 26609070]

Details:
Mitochondrial Extraction Kit was used for the preparation of mitochondrial protein extracts from brain, testes, spleen, and kidney of rats and HeLa cells.
Hornstein T, Lehmann S, Philipp D et al. Staurosporine resistance in inflammatory neutrophils is associated with the inhibition of caspase- and proteasome-mediated Mcl-1 degradation. J. Leukoc. Biol. 2015 Aug 26 [PMID: 26310832]
Kang L, Dai C, Lustig ME et al. Heterozygous SOD2 Deletion Impairs Glucose-Stimulated Insulin Secretion, but not Insulin Action in High Fat-Fed Mice. Diabetes. 2014 Jun 19 [PMID: 24947366] (WB, Mouse)

Details:
Fig 1B: Mitochondria isolated from gastrocnemius were used for WB
WB Mouse
Somasagara Ranganatha R, Hegde Mahesh, Chiruvella Kishore K et al. Extracts of strawberry fruits induce intrinsic pathway of apoptosis in breast cancer cells and inhibits tumor progression in mice. PLoS One. 2012 [PMID: 23071702] (WB) WB
Hegde Mahesh, Karki Subhas S, Thomas Elizabeth et al. Novel levamisole derivative induces extrinsic pathway of apoptosis in cancer cells and inhibits tumor progression in mice. PLoS One. 2012 [PMID: 22970136] (WB) WB
Csak T, Dolganiuc A, Kodys K et al. Mitochondrial antiviral signaling protein defect links impaired antiviral response and liver injury in steatohepatitis in mice. Hepatology. 2011 Jun [PMID: 21425308] (WB)

Details:
Cytochrome c (IMG-101A), Caspase-8 (IMG-5703), Mitochondrial extraction kit (10082k). WB: Mitochondrial and cytoplasmic extract from C57B16 MCD (methionine-choline deficient) diet fed and non MCD diet fed mice, Fig 5.
WB
Fu HY, Okada K, Liao Y et al. Ablation of C/EBP homologous protein attenuates endoplasmic reticulum-mediated apoptosis and cardiac dysfunction induced by pressure overload. Circulation. 2010 Jul 27 [PMID: 20625112] (Mouse)

Details:
Mitochondrial extraction kit (10082k). Mitochondrial fractions from mouse heart were subject to western blot using Bcl2 and Bax antibodies, Fig 4A.
Mouse
Cornille E, Abou-Hamdan M, Khrestchatisky M et al. Enhancement of L-3-hydroxybutyryl-CoA dehydrogenase activity and circulating ketone body levels by pantethine. Relevance to dopaminergic injury. BMC Neurosci. 2010 Apr 23 [PMID: 20416081]

Details:
Mitochondrial extraction kit (10082k). Mitochondrial extract was taken from brain and liver tissues.
Lee S, Van Remmen H, Csete M. Sod2 overexpression preserves myoblast mitochondrial mass and function, but not muscle mass with aging. Aging Cell. 2009 Jun [PMID: 19627269]

Details:
Mitochondrial extraction kit (10082k). Mitochondrial fractions were obtained from human myoblasts.
Show All 15 Publications.

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FAQs for Mitochondrial Kit (NBP2-29448). (Showing 1 - 1 of 1 FAQs).

  1. We are looking to isolate mtDNA that is free from nuclear DNA and nuclear DNA that is free from mtDNA. Do you know if these kits (NBP2-29447, NBP2-29448) will be able to do this? We are wanting the isolated DNA to use as a positive control for a copy number experiment.
    • You would need to use both kits in order to seperate mtDNA and nDNA. I was worried about the lysis buffer contained in both kits would force the DNA to precipitate, this can occur if the buffer is very basic or alcohol based. I also double checked to make sure all the buffers contained within the kit do not have any DNAses or RNAses present. Follow the protocol for both kits until you use the mitochondrial/nuclear lysis buffer and then proceed to collect the individual supernatants. I would proceed to use our Quik ChIP Kit. The way this kit works is by precipitating the DNA and then running it through a spin column included into the kit. Then the DNA is eluded of the column and the resulting product should be purified mDNA or nDNA depending on which supernatant you used. I reccomend on using a lot of starting material for your nuclear extract as the yield tends to be low, which is typical. I also reccomend to use 18 mega ohm DI water when starting your nuclear/mitochondrial extractions. The nuclear and mitochondrial extraction kits were designed for the purpose of extracting proteins and have not been tested for this particular application.

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