Mitochondria Isolation Kit

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

Product Details

Summary
Reactivity All-NASpecies Glossary
Kit Type
Isolation Kit

Order Details

Mitochondria Isolation Kit Summary

Description
This kit can be used to isolate cytosolic and mitochondrial fractions from the same cell or tissue preparations. The procedure 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 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.
Kit Type
Isolation Kit

Applications/Dilutions

Dilutions
  • Western Blot
Publications
Read Publications using
NBP2-29448 in the following applications:

  • WB
    10 publications

Packaging, Storage & Formulations

Storage
Storage of components varies. See protocol for specific instructions.

Kit Components

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

Alternate Names for Mitochondria Isolation Kit

  • Mitochondria Extraction Kit

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 Mitochondria Kit (NBP2-29448)(22)

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

We have publications tested in 1 application: WB.


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WB
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Human
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Mouse
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Showing Publications 1 - 10 of 22. Show All 22 Publications.
Publications using NBP2-29448 Applications Species
Hwang DJ, Kwon KC, Choi DH Et al. Comparison of intrinsic exercise capacity and response to acute exercise in ICR (Institute of Cancer Research) mice derived from three different lineages Laboratory animal research [PMID: 34348800]
Hornstein T. Molecular elucidation of apoptosis resistance in neutrophils from patients with severe trauma. Thesis (WB, Human) WB Human
Dahal S, Dubey S, Raghavan SC. Homologous recombination-mediated repair of DNA double-strand breaks operates in mammalian mitochondria Cell. Mol. Life Sci. 2017 Nov 07 [PMID: 29116362] (Mouse) Mouse
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]
Chong A, Vandana J, Li G Et Al. Checkpoint Kinase 2 Controls Insulin Secretion and Glucose Homeostasis SSRN Electronic Journal Jul 23 2021
Jeong JH, Koo JH, Yook JS, et al. Neuroprotective Benefits of Exercise and MitoQ on Memory Function, Mitochondrial Dynamics, Oxidative Stress, and Neuroinflammation in D-Galactose-Induced Aging Rats Brain sciences Jan 27 2021 [PMID: 33514020]
Dietl A, Winkel I, Pietrzyk G, et al. Skeletal muscle alterations in tachycardia-induced heart failure are linked to deficient natriuretic peptide signalling and are attenuated by RAS-/NEP-inhibition PLoS ONE Dec 4 2019 [PMID: 31800630]
Koo JH, Kang EB Effects of treadmill exercise on the regulatory mechanisms of mitochondrial dynamics and oxidative stress in the brains of high-fat diet fed rats J Exerc Nutrition Biochem Mar 31 2019 [PMID: 31010272]
Show All 22 Publications.

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Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol

FAQs for Mitochondria 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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Immune Cell Metabolic Flux Influences Type I Diabetes
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Methamphetamine with HIV induces mitochondrial dysfunction and neuronal injury through oxidative stress
By Jamshed Arslan, Pharm. D., PhD. December 1 is the World AIDS Day. Despite the combination antiretroviral therapy, 10-25% of Human Immunodeficiency Virus (HIV)-positive individuals report neurocognitive impairm...  Read full blog post.

There's an autophagy for that!
By Christina Towers, PhDA critical mechanism that cells use to generate nutrients and fuel metabolism is through a process called autophagy.  This process is complex and involves over 20 different proteins, most of which are highly conserved acro...  Read full blog post.

Dynamin-related Protein 1 (DRP1) in Mitochondria and Apoptosis.
Dynamin-related Protein 1 (DRP1) is known to function in mitochondrial and peroxisomal division and mediate membrane fission through oligomerization into ring-like structure and sever the mitochondrial membrane, through a GTP hydrolysis-dependent mech...  Read full blog post.

The Importance of the COX IV Antibody to Apoptosis Research
COX IV isoform 1 is a nuclear-encoded polypeptide chain of the Cytochrome C Oxidase enzyme, located on the mitochondrial inner membrane. Owing to its widespread distribution in human and mammalian tissues, COX IV antibodies are widely used as loading ...  Read full blog post.

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