Following the protocol will result in unfixed isolated nuclei
NUC101
Nuclei Isolation Kit: Nuclei EZ Prep
sufficient for 25 nuclei preparations (~1-10×107 cells/preparation)
Synonym(s):
Rapid mammalian nuclei isolation kit
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About This Item
NACRES:
NA.32
UNSPSC Code:
12352207
Pricing and availability is not currently available.
usage
sufficient for 25 nuclei preparations (~1-10×107 cells/preparation)
Quality Segment
shelf life
1 yr at 2‑8 °C
packaging
pkg of 1 kit
application(s)
cell analysis
foreign activity
nuclease and protease, free
shipped in
wet ice
storage temp.
2-8°C
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.
Application
Suitable as a source of nuclear components, to produce nuclei for in vitro apoptosis assays, and for functional studies.
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.
Other Notes
All procedures should be carried out on ice or 2-8°C.
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| Quality Level 200 | Quality Level 200 | Quality Level 300 | Quality Level 300 |
| usage sufficient for 25 nuclei preparations (~1-10×107 cells/preparation) | usage sufficient for 15 nuclei preparations (~1-10×107 cells or 1g of tissue per preparation) | usage kit sufficient for 10 extractions (1 ml packed cell volume), kit sufficient for 100 extractions (100 μl packed cell volume) | usage - |
| shipped in wet ice | shipped in wet ice | shipped in dry ice | shipped in ambient |
| shelf life 1 yr at 2‑8 °C | shelf life - | shelf life - | shelf life - |
| foreign activity nuclease and protease, free | foreign activity nuclease and protease, free | foreign activity - | foreign activity - |
| packaging pkg of 1 kit | packaging pkg of 1 kit | packaging - | packaging pkg of -57 tests |
Storage Class
10 - Combustible liquids
flash_point_f
Not applicable
flash_point_c
Not applicable
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At the end of the protocol I will be with fixed nuclei or fresh nuclei? I need to know for the subsequent protocol that I will do
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Can this buffer be used for frozen human lung tissue?
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Freezing and thawing processes may damage many cells and nuclei, potentially leading to the loss of nuclear components or contamination of the nuclei with cytoplasmic or other cellular components.
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Can this buffer be used with ipscs and can the incubation be conducted in a thermocyler at 2-4C instead of on ice
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Yes, this kit has been referenced to work with induced pluripotent stem cells (iPSCs). Although not tested, using a thermocycler set to 2-4°C will effectively maintain the required conditions as a viable alternative to using ice for the incubation steps during the nuclei isolation process. Please see the link below to review the publication referencing use with iPSCs.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6992778/Helpful?
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What is the pH of the EZ-prep lysis buffer
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The complete formulation for the Lysis solution is given in the following reference. The lysis buffer in the kit is formulated the same, except without the Triton X-100 and the DTT.
“Analysis of nuclear RNA”, Chapter 19, in RNA Methodologies: A Laboratory Guide for Isolation and Characterization, Robert E. Farrell, Jr., Academic Press, San Diego, p. 406-437 (1998)Helpful?
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What is the formulation of the storage solution of NUC101?
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The storage solution for NUC101 (Product No. S8933) contains 2 mM MgCl2, 0.1 mM Na2EDTA, and no detergent in a Tris buffer (pH 8.0) with 30% glycerol.
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Is the Nuclei isolation Kit Nuclei EZ Prep designed for DNA or protein isolation?
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The Nuclei isolation Kit Nuclei EZ Prep is designed for the isolation of intact nuclei containing nucleic acid and protein factors.
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Can this kit be used with vertebrate cells which are not mammalian (such a avian cells)?
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Product NUC101 has been validated for use with mammalian samples. However, it has been used in many different species by independent laboratories. One study used Nuc201 with zebrafinch samples:
https://www.nature.com/articles/s41467-021-22918-2Helpful?
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For the upcoming important phase of our project, we need the Nuclei EZ Prep Nuclei Isolation Kit. Do you have alternatives to this product in your range, which we can use for our Nuclei Isolation? Or are there any remaining stocks that we can use.
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The NUC201 Nuclei Isolation Kit is an available alternative for this product. The NUC201 method includes centrifugation in a sucrose solution which offers additional protection to the nuclei, but may result in a lower yield when compared to the NUC101. Please see the link below to review the product datasheet:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/210/075/nuc201bul.pdfFor additional questions regarding this product option, we kindly ask you to navigate to the link https://www.sigmaaldrich.com/techservice, click on "Product Technical Inquires" under the Products Section with all the required information so that a member of our team can reach out to you to assist further. Thank you.
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Can Product NUC101, Nuclei Isolation Kit, be used to capture soluble nuclear proteins?
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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.
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Can I use Product NUC101, Nuclei Isolation Kit, on frozen cells?
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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.
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