Thaw nuclei stored at -70 °C in this product’s storage buffer slowly on ice or at 2–8 °C until just thawed. Mix gently by flicking the tube or gentle pipetting; avoid vortexing or rapid warming to prevent damage. After thawing, place on ice and proceed immediately with the downstream application. Minimize freeze–thaw cycles; aliquot before freezing. . Please see the link below to access a technical bulletin.
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/528/598/nuc101bul.pdf
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.
1 of 1
This Item | |||
|---|---|---|---|
| description sufficient for 25 nuclei preparations (~1-10×107 cells/preparation) | description sufficient for 15 nuclei preparations (~1-10×107 cells or 1g of tissue per preparation) | description For mammalian tissue or cultured cells | description For plant leaves |
| shipped in wet ice | shipped in wet ice | shipped in dry ice | shipped in wet ice |
| application(s) cell analysis | application(s) cell analysis | application(s) - | application(s) - |
| 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 100 extractions (100 μl packed cell volume), kit sufficient for 10 extractions (1 ml packed cell volume) | usage kit sufficient for 30 extractions (20 g of fresh or frozen leaves) |
| Quality Level 200 | Quality Level 200 | Quality Level 300 | Quality Level 200 |
| packaging pkg of 1 kit | packaging pkg of 1 kit | packaging - | packaging - |
| storage temp. 2-8°C | storage temp. 2-8°C | storage temp. −20°C | storage temp. 2-8°C |
Storage Class
10 - Combustible liquids
Flash Point (°F)
Not applicable
Flash Point (°C)
Not applicable
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Can I use nuclei prepared by this kit for CUT&TAG or CUT&RUN assay?
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How should we thaw the nuclei before use after storage at -70°C in storage buffer?
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Does the lysis buffer contain detergent? If so, is there a risk that detergent-mediated nuclear disruption could cause leakage of nuclear proteins?
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No, this product does not contain a detergent.
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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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Following the protocol will result in unfixed isolated nuclei
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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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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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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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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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