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.

Name: Mitochondria Isolation Kit
Brand: Novus Biologicals
SKU: NBP2-29448

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.

Mitochondria Isolation Kit

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-29448

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Product Summary
Scientific Data
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Background
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Key Product Details

Species

Non-species specific

Applications

Western Blot

Kit Type

Isolation Kit

Product Summary for Mitochondria Isolation Kit

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.

Scientific Data Images for Mitochondria Isolation Kit

Western Blot: Mitochondria Isolation Kit [NBP2-29448]

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 antibody (loading control). Isolation kit efficiently separated the two fractions: No Cytochrome C (specific to mitochondria) was detected in the cytosolic extract.

Kit Contents for Mitochondria Isolation Kit

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

Formulation, Preparation, and Storage

Concentration

Concentration is not relevant for this product. Please see the protocols for proper use of this product.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Storage

Storage of components varies. See protocol for specific instructions.

Background: Mitochondria

NB600-556 recognizes a 60kDa non-glycosylated protein component of mitochondria found in human cells. It can be used to stain the mitochondria in cell or tissue preparations and can be used as a marker of the mitochondria in subcellular fractions. This monoclonal antibody is part of a new panel of reagents which recognizes subcellular organelles or compartments of human cells. These markers may be useful in identification of these organelles in cells, tissues, and biochemical preparations. MAb-2 produces a spaghetti-like staining pattern in the cytoplasm of normal and malignant human cells and may be used to stain the mitochondria in paraformaldehyde fixed frozen and formalin-fixed, paraffin-embedded tissues.

Alternate Names

Mitochondria Extraction Kit

Additional Mitochondria Products

Product Documents for Mitochondria Isolation Kit

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Protocols & Manuals

Download Manual - Mitochondria Isolation Kit

COA

Product Specific Notices for Mitochondria Isolation Kit

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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