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: Nuclear Extraction Kit
Brand: Novus Biologicals
SKU: NBP2-29447

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.

Nuclear Extraction Kit

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-29447

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Key Product Details

Applications

Western Blot

Kit Type

Extraction Kit

Product Summary for Nuclear Extraction Kit

The Nuclear Extraction Kit provides a simple and convenient method for the isolation of nuclear and cytoplasmic extracts from mammalian cells and tissue samples. This procedure is relevant to the monitoring of translocation of cell signaling molecules from cytoplasm to the nucleus. Examples include translocation of NF-kB molecules to the nucleus in TNF-alpha treated cells, and translocation of mitogen-activated protein kinase to the nucleus in growth factor treated cells.

The Nuclear Extraction Kit can be used in the preparation of purified proteins for use in Western blotting, Eletrophoretic Mobility Assays (EMSA) and preparative purification of nuclear proteins.

Scientific Data Images for Nuclear Extraction Kit

Western Blot: Nuclear Extraction Kit [NBP2-29447]

Western Blot: Nuclear Extraction Kit [NBP2-29447] - 20 ug of HeLa cell proteins separated on 4-20% SDS-PAGE. Lane 1: Nuclear extract. Lane 2: Cytoplasmic extract. Lane 3: Total cell lysate.

Kit Contents for Nuclear Extraction Kit

10% Detergent Solution (10 mL)
100 mM PMSF (10 mL)
100X Protease Inhibitor Cocktail - PIC (100 uL)
10X Hypotonic Lysis Buffer (10 mL)
10X PBS (2 x 50 mL)
1M DTT for nuclear extraction from tissue (100 uL)
1X Nuclear Extraction Buffer (10 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: Nuclear

Nuclear Extraction

Alternate Names

Nuclear Extraction, Nuclear Extraction Kit

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Protocols & Manuals

Download Manual - Nuclear Extraction Kit

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Product Specific Notices for Nuclear Extraction 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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