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NXTRACT

Sigma-Aldrich

NuCLEAR™ Extraction Kit

For mammalian tissue or cultured cells

Synonym(s):

Nuclear Isolation Kit

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About This Item

UNSPSC Code:
41105517
NACRES:
NA.56

Quality Level

300

usage

 kit sufficient for 10 extractions (1 ml packed cell volume)
 kit sufficient for 100 extractions (100 μl packed cell volume)

technique(s)

protein extraction: suitable
western blot: suitable

shipped in

dry ice

storage temp.

−20°C

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.

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.

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.
A number of different procedures in the detailed technical bulletin enable the selection that best fits a particular application. For example, choose between detergent and non-detergent extraction of nuclear protein or between the standard hypotonic lysis buffer for most cell types and isotonic lysis buffer for fragile cells. In addition, the kit provides a procedure for salt reduction from the nuclear extract with dilution buffer. NuCLEAR offers the flexiblity you need for optimal protein extraction. Extracts can be prepared in less than 2 hours and are highly pure since there is little or no cross-contamination between nuclear and cytoplasmic extracts.

Linkage

Recommended Antibodies for Immunodetection L1293, N2662, AMAB90549

Legal Information

NuCLEAR is a trademark of Sigma-Aldrich Co. LLC

Kit Components Also Available Separately

Product No.
Description
SDS
  • 3× Dilution and Equilibration Buffer 90 mL
  • P8340Protease Inhibitor Cocktail 1 mLSDS

Pictograms

Corrosion
GHS05

Signal Word

Danger

Hazard Statements

H290,H314

Hazard Classifications

Eye Dam. 1 - Met. Corr. 1 - Skin Corr. 1A

Storage Class Code

8A - Combustible corrosive hazardous materials

Flash Point(F)

188.6 °F - closed cup

Flash Point(C)

87 °C - closed cup

Regulatory Information

常规特殊物品

Choose from one of the most recent versions:

Certificates of Analysis (COA)

Lot/Batch Number
0000149724
0000149724
0000114896
059M4108V
11181117

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If you require a particular version, you can look up a specific certificate by the Lot or Batch number.

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  1. 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.

  2. 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.

  3. 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).

  4. 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.

  5. 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.

  6. 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.

  7. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

  8. 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.

  9. 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. 

  10. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

Qing Wang et al.
American journal of physiology. Regulatory, integrative and comparative physiology, 302(9), R1025-R1033 (2012-03-10)
We previously reported that excess of deoxycorticosterone-acetate (DOCA)/salt-induced cardiac hypertrophy in the absence of hypertension in one-renin gene mice. This model allows us to study molecular mechanisms of high-salt intake in the development of cardiovascular remodeling, independently of blood pressure
K A Lee et al.
Gene analysis techniques, 5(2), 22-31 (1988-03-01)
A convenient and rapid method for preparing soluble extracts from the nuclei of as few as 3 x 10(7) mammalian cells (miniextract procedure) is described. By several criteria, miniextracts are comparable to nuclear extracts prepared from large numbers of cells
Isolation of intact nuclei for nuclear extract preparation from a fragile B-lymphocyte cell line.
R B Dyer et al.
BioTechniques, 19(2), 192-195 (1995-08-01)
Fangyuan Dong et al.
EBioMedicine, 39, 472-483 (2018-12-12)
Accumulating evidence has revealed the pivotal role of epigenetic regulation in the pathogenesis of liver disease. However, the epigenetic mechanism that accounts for hepatic stellate cells (HSCs) activation in liver fibrosis remains largely unknown. Primary HSCs were used to screen
F Guidez et al.
Molecular and cellular biology, 18(7), 3851-3861 (1998-06-25)
Granulocyte-macrophage colony-stimulating factor (GM-CSF) and macrophage colony-stimulating factor (M-CSF) independently stimulate the proliferation and differentiation of macrophages from bone marrow progenitor cells. Although the GM-CSF and M-CSF receptors are unrelated, both couple to Ras-dependent signal transduction pathways, suggesting that these
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