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)
Sinónimos:
Rapid mammalian nuclei isolation kit
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| Talla/SKU | Disponibilidad | Precio |
|---|---|---|
1 kit | Comprobar disponibilidad del carrito | $327.000 |
Acerca de este artículo
NACRES:
NA.32
UNSPSC Code:
12352207
$327.000
Comprobar disponibilidad del carrito
Servicio técnico
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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
Este artículo | |||
|---|---|---|---|
| description sufficient for 25 nuclei preparations (~1-10×107 cells/preparation) | description For mammalian tissue or cultured cells | description BioUltra, Molecular Biology | description Far red nuclear staining dye with greater photostability than traditional blue fluorescent nuclear stains such as DAPI and Hoechst 33342. |
| packaging pkg of 1 kit | packaging - | packaging - | packaging - |
| shipped in wet ice | shipped in dry ice | shipped in - | shipped in ambient |
| storage temp. 2-8°C | storage temp. −20°C | storage temp. 2-8°C | storage temp. 2-8°C |
| foreign activity nuclease and protease, free | foreign activity - | foreign activity - | foreign activity - |
| shelf life 1 yr at 2‑8 °C | shelf life - | shelf life - | shelf life - |
| usage sufficient for 25 nuclei preparations (~1-10×107 cells/preparation) | usage kit sufficient for 10 extractions (1 ml packed cell volume), kit sufficient for 100 extractions (100 μl packed cell volume) | usage - | usage - |
Clase de almacenamiento
10 - Combustible liquids
Punto de inflamación (°F)
Not applicable
Punto de inflamación (°C)
Not applicable
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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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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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