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NUC101 Sigma

Nuclei Isolation Kit: Nuclei EZ Prep

sufficient for 25 nuclei preparations (~1-10×107 cells/preparation)

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Description

Frequently Asked Questions

Frequently Asked Questions are available for this Product.

Application

Suitable as a source of nuclear components, to produce nuclei for in vitro apoptosis assays, and for functional studies.

Other Notes

All procedures should be carried out on ice or 2-8°C.

Biochem/physiol Actions

The protocol provides a high yield of nuclei from commonly used mammalian cells, including both adherent (e.g., HEK293 and COS7) and non-adherent (e.g., Jurkat and HFN7.1) cell lines and peripheral blood mononuclear cells (PBMCs). The preparations are suitable for many cell biology applications, e.g., as a source of nuclear components such as chromatin, genomic DNA, histones and nuclear RNA/RNP, produces nuclei for in vitro apoptosis assays, and functional studies such as examination of the transcriptional status of cells.

General description

The Nuclei EZ Prep Kit was developed for the rapid isolation of nuclei from mammalian cells. The kit provides a high yield of nuclei from commonly used cell types.

Price and Availability


Ultrafree Centrifugal Filter Units

All labs need water

Standard component

Description

Product #

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Nuclei EZ Lysis Buffer 200 mL SDS N3408
Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport

Frequently Asked Questions

Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis. These documents are located on the product detail page under Useful Links & Tools. Click on the following link to search for a Certificate of Analysis. Please click the following link to see the details on our Product Dating Information.
How do I get lot-specific information or a Certificate of Analysis?
A Certificate of Analysis is available by lot number and can be obtained through our Advanced Search Option: http://www.sigmaaldrich.com/catalog/AdvancedSearchPage.do
Can Product NUC101, Nuclei Isolation Kit, be used to capture soluble nuclear proteins?
The kit was originally designed to produce nuclei that are competent for transcription run-on/run-off assays to measure transcription from pre-engaged polymerase complexes on the nuclear DNA. It was not designed for the separation or isolation of soluble nuclear proteins. The researcher might be advised to try the NXTRACT nuclear extraction kit, which allows for the isolation of "intact" nuclei from other cellular components before the lysis of the nuclear membrane.  For information on the NXTRACT kit, follow this link:  http://www.sigmaaldrich.com/catalog/search/ProductDetail/SIGMA/NXTRACT
Can I use Product NUC101, Nuclei Isolation Kit, on frozen cells?
It is likely that the freezing and thawing processes will damage many of the cells and nuclei, which could lead to loss of nuclear components and/or contamination of the nuclei with cytoplasmic or other cellular components.
Should I use trypsin to harvest my adherent cells before using Product NUC101, Nuclei Isolation Kit?
In general, this is not advised. We have never tried detaching the cells first with trypsin. We would not recommend it, because residual trypsin could clip nuclear proteins and cause problems in downstream applications.
How do I find price and availability?
There are several ways to find pricing and availability for our products.  Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers. 
What is the Department of Transportation shipping information for this product?
Transportation information can be found in Section 14 of the product's (M)SDS. To access the shipping information for this material, use the link on the product detail page for the product, or search here. 
My question is not addressed here, how can I contact Technical Service for assistance?
Use the option to the right to "Ask a Question" by email of a Technical Service Scientist.
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Protocols & Articles

Articles

Organelle Isolation | BioFiles Volume 6, Number 5 — Centrifugation

The isolation of subcellular fractions by centrifugation is a commonly used technique and is widely applicable across multiple cell and tissue types. Because organelles differ in their size, shape, a...
Mark Frei
BioFiles v6 n5, 22–25
Keywords: Capture ELISA, Cell culture, Centrifugation, Diffusion, Direct immunofluorescence, Dot blot, Enzyme-linked immunosorbent assay, Flow cytometry, Immunoassay, Immunocytochemistry, Immunoelectrophoresis, Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Indirect ELISA, Individual protein Immunoprecipitation, Microarray Analysis, Microscopy, Radioimmunoassay, Scanning electron microscopy, Western blot

Peer-Reviewed Papers
15

References

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