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.
Read Publications using NBP2-29447 in the following applications:
FAQs for Nuclear Kit (NBP2-29447). (Showing 1 - 1 of 1 FAQs).
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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