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15338
CUTRUN Antibody Binding Buffer
CUT&RUN Kits & Reagents
Buffer

CUT&RUN Antibody Binding Buffer #15338

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CUT and RUN Image 1: CUTRUN Antibody Binding Buffer

Product Usage Information

For the CUT&RUN assay, we recommend adding 1 μl 100X Spermidine #27287, 0.5 μl Protease Inhibitor Cocktail (200X) #7012, and 2.5 μl Digitonin Solution #16359 to 96 μl CUT&RUN Antibody Binding Buffer (100 μl per reaction) right before use. Please keep on ice during use. The volume of Digitonin Solution #16359 in the CUT&RUN Antibody Binding Buffer can be adjusted based on the specific cell line.

Storage

Store CUT&RUN Antibody Binding Buffer at 4°C. This product is stable for at least 12 months.

Product Description

The CUT&RUN Antibody Binding Buffer provides enough reagent to support 48 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. An appropriate amount of Digitonin Solution #16359, 100X Spermidine #27287, and Protease Inhibitor Cocktail (200X) #7012 should be added to this product right before use. Please keep on ice during use.

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

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

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 # 15338
Cat. # Size Qty. Price
15338L
5 ml  (2 x 2.5 ml) $ 60