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Merck
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주요 문서

NUC201

Sigma-Aldrich

Nuclei Isolation Kit: Nuclei PURE Prep

sufficient for 15 nuclei preparations (~1-10×107 cells or 1g of tissue per preparation)

동의어(들):

Sucrose centrifugation nuclei isolation

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크기 선택

1 KIT
₩1,168,062

₩1,168,062


재고 있음세부사항



크기 선택

보기 변경
1 KIT
₩1,168,062

About This Item

UNSPSC 코드:
12352207
NACRES:
NA.32

₩1,168,062


재고 있음세부사항


사용

sufficient for 15 nuclei preparations (~1-10×107 cells or 1g of tissue per preparation)

Quality Level

포장

pkg of 1 kit

저장 조건

dry at room temperature

응용 분야

cell analysis

외래 활성

nuclease and protease, free

배송 상태

wet ice

저장 온도

2-8°C

애플리케이션

For preparation of pure nuclei and fragile nuclei from cell lines and solid tissues.

생화학적/생리학적 작용

The protocol incorporates centrifugation through a dense sucrose cushion to protect nuclei and strip away cytoplasmic contaminants. High yield has been obtained from common cell lines (Jurkat, HFN7.1, COS7, HEK293 and MDCK) and tissues (spleen and liver). These 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.
The protocol incorporates centrifugation through a dense sucrose cushion to protect nuclei and strip away cytoplasmic contaminants. The sucrose concentration that is suitable for a particular cell type is determined empirically by the user. The sucrose concentrate and sucrose cushion buffer give the user flexibility to modify the density of the sucrose cushion as appropriate. High yield has been obtained from common cell lines (Jurkat, HFN7.1, COS7, HEK293 and MDCK) and tissues (spleen and liver). These 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.

키트 구성품 전용

제품 번호
설명

  • Nuclei PURE Lysis Buffer 180 mL

관련 제품

제품 번호
설명
가격

픽토그램

CorrosionEnvironment

신호어

Danger

유해 및 위험 성명서

Hazard Classifications

Aquatic Acute 1 - Aquatic Chronic 2 - Eye Dam. 1 - Skin Irrit. 2

Storage Class Code

10 - Combustible liquids

Flash Point (°F)

Not applicable

Flash Point (°C)

Not applicable


가장 최신 버전 중 하나를 선택하세요:

시험 성적서(COA)

Lot/Batch Number

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특정 버전이 필요한 경우 로트 번호나 배치 번호로 특정 인증서를 찾을 수 있습니다.

이 제품을 이미 가지고 계십니까?

문서 라이브러리에서 최근에 구매한 제품에 대한 문서를 찾아보세요.

문서 라이브러리 방문

Analysis of nuclear RNA, Chapter 14.
Robert E. Farrell, Jr., ed.
RNA Methodologies: A Laboratory Guide for Isolation and Characterization, 235-263 (1993)
Nuclei from rat liver: isolation method that combines purity with high yield.
G Blobel et al.
Science (New York, N.Y.), 154(3757), 1662-1665 (1966-12-30)
Tyler G Ekins et al.
eLife, 9 (2020-11-06)
Type I lissencephaly is a neuronal migration disorder caused by haploinsuffiency of the PAFAH1B1 (mouse: Pafah1b1) gene and is characterized by brain malformation, developmental delays, and epilepsy. Here, we investigate the impact of Pafah1b1 mutation on the cellular migration, morphophysiology
Michaela Patterson et al.
Nature genetics, 49(9), 1346-1353 (2017-08-08)
Adult mammalian cardiomyocyte regeneration after injury is thought to be minimal. Mononuclear diploid cardiomyocytes (MNDCMs), a relatively small subpopulation in the adult heart, may account for the observed degree of regeneration, but this has not been tested. We surveyed 120
Sundong Lin et al.
Diabetes & metabolism journal, 44(1), 158-172 (2019-11-09)
Epithelial-to-mesenchymal transition (EMT) is required for renal fibrosis, which is a characteristic of diabetic nephropathy (DN). Our previous study demonstrated that fibroblast growth factor 21 (FGF21) prevented DN associated with the suppressing renal connective tissue growth factor expression, a key

문서

Centrifugation separates organelles based on size, shape, and density, facilitating subcellular fractionation across various samples.

질문

1–8 / 8 질문  
  1. What is the expected loss (%) of nuclei due resulting from the sucrose density gradient purification step? I am planning an experiment with small tissue masses (50-100mg, ~1x10^6 nuclei) and would like to optimise yield as much as possible. 

    1 답변
    1. Yield will vary between cell lines and sample types, so the expected loss must be determined empirically for each sample. The technical bulletin notes that typical nuclei yields are greater than 30%, but some cell lines and sample types may experience lower yields.

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  2. Is the kit compatible to frozen tissue stored in RNAlater?

    1 답변
    1. This kit has not been evaluated for use with frozen samples. It is highly recommended to use fresh tissue samples with product. The freezing and thawing processes could potentially damage many of the cells and nuclei, leading to the loss of functionality/stability of some nuclear molecules/entities.

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  3. Is there EDTA present in buffer L9286? Additionally, is the lysis buffer L9286 hypotonic or isotonic/hypertonic?

    1 답변
    1. The lysis buffer is an isoosmotic sucrose buffer and it does contain EDTA.

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  4. Is it possible to split the sample in half if the right size tubes for one of the steps are not available?

    1 답변
    1. As per internal notes, the kit can be scaled down to as small as 1.5 ml tubes, provided that the centrifugation speed and time are not deviated from. For example, the sucrose cushion centrifugation step must be carried out for 45 minutes at 30,000 × g, preferably in an ultracentrifuge swinging rotor, as recommended in the Technical Bulletin.

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  5. Would the nuclei isolation protocol need to be modified when using NUC201 as an alternative to NUC101-1kt?

    1 답변
    1. The NUC201 and NUC101 kits are very different. NUC101 has two components—a different lysis buffer and a storage buffer. In contrast, NUC201 has a different lysis buffer and a total of 5 kit components. Additionally, DTT needs to be used, although it is listed as a required but not provided reagent. Instructions for Use will be included with each kit. If the procedure is not provided with the kit, a PDF of the kit instructions can be downloaded from the product detail pages for each kit. Due to the significant procedural differences, it is important to use the specific Instructions for Use for each kit.

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  6. Is it possible to start the nuclei isolation process from animal tissue or cells by using a frozen sample or cell pellet (kept in -80°C)?

    1 답변
    1. The freezing and thawing processes could potentially damage many of the cells and nuclei, leading to the loss of nuclear components and possible contamination of the nuclei with cytoplasmic or other cellular components. Testing on NUC201 has been conducted on fresh tissue.

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  7. Is it possible to start with frozen tissue?

    1 답변
    1. It is probable that the freezing and thawing processes may harm a significant number of cells and nuclei, potentially resulting in the loss of nuclear components and/or contamination of the nuclei with cytoplasmic or other cellular components.

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  8. Is the kit compatible with Tris or DTT, as our internal protocol is not compatible with these components?

    1 답변
    1. The NUC201 kit is compatible with Tris as well as DTT. None of the components in the kit, such as sucrose, Triton X100, and glycerin, are known to be incompatible with Tris. Additionally, as these components do not contain sulfur, there is no reason to expect any incompatibility with DTT.

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