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
Fortfahren mit
NUC101
Zellkernisolierung - Kit: Nuclei EZ Prep
sufficient for 25 nuclei preparations (~1-10×107 cells/preparation)
Synonym(e):
Rapid mammalian nuclei isolation kit
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| Größe/SKU | Verfügbarkeit | Preis |
|---|---|---|
1 kit | Warenkorb auf Verfügbarkeit prüfen | € 434,00 |
Über diesen Artikel
NACRES:
NA.32
UNSPSC Code:
12352207
€ 434,00
Warenkorb auf Verfügbarkeit prüfen
Technischer Dienst
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Unterstützung erhaltenusage
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
Das Nuclei EZ Prep-Kit wurde speziell für die rasche Isolierung der Zellkerne aus Säugetierzellen entwickelt. Das Kit liefert einen hohen Ertrag an Zellkernen aus häufig verwendeten Zelltypen.
Application
Geeignete Quelle für Nuklearkomponenten zur Produktion von Zellkernen für In-vitro-Apoptose-Assays und für funktionelle Studien.
Biochem/physiol Actions
Das Protokoll liefert einen hohen Ertrag an häufig verwendeten Säugetierzellkernen, sowohl von adhärenten (z. B. HEK293 und COS7) als auch nicht adhärenten (z. B. Jurkat und HFN7.1) Zelllinien und peripheren mononuklearen Blutzellen (PBMC). Die Präparationen eignen sich für eine Vielzahl von Zellbiologie-Anwendungen, z. B. als Quelle für Nuklearkomponenten wie Chromatin, genomische DNA, Histone und nukleäre RNA/RNP, die Produktion von Zellkernen für In-vitro-Apoptose-Assays und funktionelle Studien, wie die Untersuchung des Transkriptionsstatus von Zellen.
Other Notes
Alle Verfahren sollten auf Eis oder bei 2 - 8 °C durchgeführt werden.
1 of 1
Dieser Artikel | |||
|---|---|---|---|
| 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 |
Lagerklasse
10 - Combustible liquids
Flammpunkt (°F)
Not applicable
Flammpunkt (°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?
1 answer-
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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
1 answer-
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)?
1 answer-
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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