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NXTRACT Sigma

CelLytic NuCLEAR Extraction Kit

For mammalian tissue or cultured cells

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Description

Frequently Asked Questions

Frequently Asked Questions are available for this Product.

Other Notes

Within this kit is a complete system for preparing nuclear and cytoplasmic protein extracts from mammalian tissue or cultured cells. All reagents necessary for extraction are included.

Linkage

Recommended Antibodies for Immunodetection L1293, N2662, AMAB90549

Application

NXTRACT kit was used to study the impact of salt on cardiac differential gene expression and coronary lesion in normotensive mineralocorticoid-treated mice. It was also used to test the therapeutic potential of andrographolide for treating endometriosis.

General description

The procedure for the nuclear protein extraction method is to allow cells to swell with hypotonic buffer. The cells are then disrupted, the cytoplasmic fraction is removed, and the nuclear proteins are released from the nuclei by a high salt buffer.

Legal Information

CelLytic is a trademark of Sigma-Aldrich Co. LLC

NuCLEAR is a trademark of Sigma-Aldrich Co. LLC

Price and Availability


Amplified Detection

Ultrafree Centrifugal Filter Units

All labs need water

Kit component only

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10× Lysis Buffer, Hypotonic 7 mL SDS    
3× Dilution and Equilibration Buffer 90 mL SDS    
1 M DTT .4 mL SDS    
Extraction Buffer 10 mL SDS    
IGEPAL® CA-630 10% Solution 4 mL SDS    
5× Lysis Buffer, Isotonic 14 mL SDS    

Standard component

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Protease Inhibitor Cocktail 1 mL SDS P8340
Safety & Documentation

Safety Information

Symbol 
GHS05  GHS05
Signal word 
Danger
Hazard statements 
Precautionary statements 
RIDADR 
UN 3316 9
Flash Point(F) 
188.6 °F
Flash Point(C) 
87 °C

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.
What are the options for lysis of my cells to make extracts when using Product NXTRACT, CelLytic™ NuCLEAR™ Extraction Kit?
The use of detergent will lead to maximum protein yield. However we find that the hypotonic lysis is extremely effective and in addition for very fragile cells we offer an isotonic lysis option.
Can Product NXTRACT, CelLytic™ NuCLEAR™ Extraction Kit, be used to make nuclear protein extracts for EMSAs/DNA binding studies?
Several references describe the use of the N-XTRACT kit in sample preparation for EMSA work.1. Xie, J., et al., β2-Microglobulin as a negative regulator of the immune system: high concentrations of the protein inhibit in vitro generation of functional dendritic cells. Blood, 101(10), 4005-4012 (2003).2. Marambaud, P., et al., A CBP Binding Transcriptional Repressor Produced by the PS1/ε-Cleavage of N-Cadherin Is Inhibitedby PS1 FAD Mutations. Cell, 114, 635-645 (2003).3. Chen, G., et al., Phosphorylated FADD induces NF-κB, perturbs cell cycle, and is associated with poor outcome in lung adenocarcinomas. Proc. Nat. Acad. Sci. USA, 102(35), 12507-12512 (2005).
Is Product NXTRACT, CelLytic™ NuCLEAR™ Extraction Kit, suitable for use on tissues? 
We have used the kit with several tissues: Mouse lung and brain, Rat liver and Rabbit muscle. For these tissues we usually used the kit with 100 mg tissue. After extraction with 140 μl Extraction buffer, we obtained an average yield of protein concentration as follows: 3.2 mg/ml for brain, 4.1 mg/ml for lung, 3.3 mg/ml for muscle and 10 mg/ml for liver.
When using Product NXTRACT, CelLytic™ NuCLEAR™ Extraction Kit, what can I use for the mechanical lysis of my cells?
We recommend the use of a glass tissue homogenizer, with a type B pestle, or alternatively a syringe with a narrow-gauge (No. 27) hypodermic needle may be used.  To view this information on the NXTRACT Technical Bulletin, click on the following link:  http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Bulletin/nxtractbul.Par.0001.File.tmp/nxtractbul.pdf
How "pure" is the nuclear extraction using Product NXTRACT, CelLytic™ NuCLEAR™ Extraction Kit? 
It is important to keep in mind that this kit will provide a crude nuclear fraction.  Depending on the care taken at the step where the nuclei are pelleted and the supernatant containing the cytoplasmic fraction is removed, we have found that the cytoplasmic contamination of the nuclear pellet may be a few percent. 
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
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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