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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1 KIT
MXP 8,606.00
MXP 8,606.00
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About This Item
NACRES:
NA.32
UNSPSC Code:
12352207
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usage
sufficient for 25 nuclei preparations (~1-10×107 cells/preparation)
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
Quality Level
1 of 3
This Item | ||
|---|---|---|
| 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) |
| shelf life 1 yr at 2‑8 °C | shelf life - | shelf life - |
| packaging pkg of 1 kit | packaging pkg of 1 kit | packaging - |
| application(s) cell analysis | application(s) cell analysis | application(s) - |
| foreign activity nuclease and protease, free | foreign activity nuclease and protease, free | foreign activity - |
| shipped in wet ice | shipped in wet ice | shipped in dry ice |
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.
related product
Product No.
Description
Pricing
Storage Class
10 - Combustible liquids
flash_point_f
Not applicable
flash_point_c
Not applicable
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Christina N Vallianatos et al.
Communications biology, 3(1), 278-278 (2020-06-03)
Histone H3 lysine 4 methylation (H3K4me) is extensively regulated by numerous writer and eraser enzymes in mammals. Nine H3K4me enzymes are associated with neurodevelopmental disorders to date, indicating their important roles in the brain. However, interplay among H3K4me enzymes during
Analysis of nuclear RNA, Chapter 14.
Robert E. Farrell, Jr., ed.
RNA Methodologies: A Laboratory Guide for Isolation and Characterization, 235-263 (1993)
Yu-Yin Shih et al.
PloS one, 6(10), e26236-e26236 (2011-10-25)
Retinoic acid (RA) has been approved for the differentiation therapy of neuroblastoma (NB). Previous work revealed a correlation between glucose-regulated protein 75 (GRP75) and the RA-elicited neuronal differentiation of NB cells. The present study further demonstrated that GRP75 translocates into
Buqing Ye et al.
Nature communications, 8(1), 1518-1518 (2017-11-16)
Lymphoid lineage commitment is an important process in haematopoiesis, which forms the immune system to protect the host from pathogen invasion. However, how multipotent progenitors (MPP) switch into common lymphoid progenitors (CLP) or common myeloid progenitors (CMP) during this process
Naomi Habib et al.
Nature methods, 14(10), 955-958 (2017-08-29)
Single-nucleus RNA sequencing (sNuc-seq) profiles RNA from tissues that are preserved or cannot be dissociated, but it does not provide high throughput. Here, we develop DroNc-seq: massively parallel sNuc-seq with droplet technology. We profile 39,111 nuclei from mouse and human
Articles
Centrifugation separates organelles based on size, shape, and density, facilitating subcellular fractionation across various samples.
Related Content
Instructions
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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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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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What is the Department of Transportation shipping information for this product?
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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.
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Should I use trypsin to harvest my adherent cells before using Product NUC101, Nuclei Isolation Kit?
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In general, this is not advised. We have never tried detaching the cells first with trypsin. We would not recommend it, because residual trypsin could clip nuclear proteins and cause problems in downstream applications.
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