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NUC201

Sigma-Aldrich

Nuclei Isolation Kit: Nuclei PURE Prep

sufficient for 15 nuclei preparations (~1-10×107 cells or 1g of tissue per preparation)

Synonym(s):

Sucrose centrifugation nuclei isolation

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About This Item

UNSPSC Code:
12352207
NACRES:
NA.32

usage

sufficient for 15 nuclei preparations (~1-10×107 cells or 1g of tissue per preparation)

Quality Level

packaging

pkg of 1 kit

storage condition

dry at room temperature

application(s)

cell analysis

foreign activity

nuclease and protease, free

shipped in

wet ice

storage temp.

2-8°C

Application

For preparation of pure nuclei and fragile nuclei from cell lines and solid tissues.

Biochem/physiol Actions

The protocol incorporates centrifugation through a dense sucrose cushion to protect nuclei and strip away cytoplasmic contaminants. High yield has been obtained from common cell lines (Jurkat, HFN7.1, COS7, HEK293 and MDCK) and tissues (spleen and liver). These 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.
The protocol incorporates centrifugation through a dense sucrose cushion to protect nuclei and strip away cytoplasmic contaminants. The sucrose concentration that is suitable for a particular cell type is determined empirically by the user. The sucrose concentrate and sucrose cushion buffer give the user flexibility to modify the density of the sucrose cushion as appropriate. High yield has been obtained from common cell lines (Jurkat, HFN7.1, COS7, HEK293 and MDCK) and tissues (spleen and liver). These 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.

Kit Components Only

Product No.
Description

  • Nuclei PURE Lysis Buffer 180 mL

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CorrosionEnvironment

Signal Word

Danger

Hazard Statements

Hazard Classifications

Aquatic Acute 1 - Aquatic Chronic 2 - Eye Dam. 1 - Skin Irrit. 2

Storage Class Code

10 - Combustible liquids

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Regulatory Listings

Regulatory Listings are mainly provided for chemical products. Only limited information can be provided here for non-chemical products. No entry means none of the components are listed. It is the user’s obligation to ensure the safe and legal use of the product.

PDSCL

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PRTR

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FSL

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ISHL Indicated Name

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ISHL Notified Names

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Cartagena Act

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JAN Code

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Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Nuclei from rat liver: isolation method that combines purity with high yield.
G Blobel et al.
Science (New York, N.Y.), 154(3757), 1662-1665 (1966-12-30)
Wei Chen et al.
The Journal of biological chemistry, 283(24), 16293-16298 (2008-04-10)
Previous studies have shown that the acute stimulation of endothelial nitric-oxide synthase (eNOS) mRNA transcription by laminar shear stress is dependent on nuclear factor kappa B (NFkappaB) subunits p50 and p65 binding to a shear stress response element (SSRE) in
Analysis of nuclear RNA, Chapter 14.
Robert E. Farrell, Jr., ed.
RNA Methodologies: A Laboratory Guide for Isolation and Characterization, 235-263 (1993)

Articles

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, and density, centrifugation can be easily employed to separate and purify organelle fractions from gently homogenized samples.

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