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48105
CUTRUN 4X Stop Buffer
CUT&RUN Kits & Reagents
Buffer

CUT&RUN 4X Stop Buffer #48105

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CUT and RUN Image 1: CUTRUN 4X Stop Buffer

Product Usage Information

For the CUT&RUN assay, we recommend adding 37.5 μl CUT&RUN 4X Stop Buffer, 3.75 μl Digitonin Solution #16359, and 0.75 μl RNAse A (10 mg/ml) #7013 to 108 μl nuclease-free water (150 μl per reaction) right before use. Then incubate with the sample at 37°C for 10 min. An appropriate amount of Sample Normalization Spike-In DNA can be added to CUT&RUN 4X Stop Buffer, if desired.

Storage

Store CUT&RUN 4X Stop Buffer at -20°C. This product is stable for at least 12 months.

Product Description

The CUT&RUN 4X Stop Buffer provides enough reagent to support 24 CUT&RUN assays. This product is formulated for optimal performance in the CUT&RUN assay and each lot is tested and validated using the CUT&RUN Assay Kit #86652. This product should be diluted to 1X using nuclease-free water and an appropriate amount of Digitonin Solution #16359 and RNAse A (10 mg/ml) #7013 should be added right before use. Addition of Spike-In DNA is optional.

Background

Like the chromatin immunoprecipitation (ChIP) assay, Cleavage Under Targets and Release Using Nuclease (CUT&RUN) is a powerful and versatile technique used for probing protein-DNA interactions within the natural chromatin context of the cell (1-4). CUT&RUN provides a rapid, robust, and true low cell number assay for detection of protein-DNA interactions in the cell. Unlike the ChIP assay, CUT&RUN is free from formaldehyde cross-linking, chromatin fragmentation, and immunoprecipitation, making it a much faster and more efficient method for enriching protein-DNA interactions and identifying target genes. CUT&RUN can be performed in less than one day, from live cells to purified DNA, and has been shown to work with as few as 500-1,000 cells per assay (1,2). Instead of fragmenting all of the cellular chromatin as done in ChIP, CUT&RUN utilizes an antibody-targeted digestion of chromatin, resulting in much lower background signal than seen in the ChIP assay. As a result, CUT&RUN requires only 1/10th the sequencing depth that is required for ChIP-seq assays (1,2). Finally, the inclusion of simple spike-in control DNA allows for accurate quantification and normalization of target-protein binding that is not possible with the ChIP method. This provides for effective normalization of signal between samples and between experiments.

  1. Skene, P.J. and Henikoff, S. (2017) Elife 6, pii: e21856. doi: 10.7554/eLife.21856.
  2. Skene, P.J. et al. (2018) Nat Protoc 13, 1006-19.
  3. Meers, M.P. et al. (2019) Elife 8, pii: e46314. doi: 10.7554/eLife.46314.
  4. Meers, M.P. et al. (2019) Mol Cell 75, 562-575.e5.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
To Purchase # 48105S
Product # Size Price
48105S
1 ml $ 20