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16359
Digitonin Solution
CUT&RUN Kits & Reagents
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Digitonin Solution #16359

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CUT and RUN Image 1: Digitonin Solution
CUT&RUN and ChIP assays were performed with HCT 116 cells and Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb #9751. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. Panel A compares enrichment of H3K4me3 across chromosome 12 (upper), while Panel B compares enrichment at the GAPDH gene (lower), a known target of H3K4me3. The input tracks are from the CUT&RUN input sample.
CUT and RUN Image 2: Digitonin Solution
CUT&RUN and ChIP assays were performed with HeLa cells and Rpb1 CTD (4H8) Mouse mAb #2629. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. Panel A compares enrichment of Rpb1 across chromosome 12 (upper), while Panel B compares enrichment at the GAPDH gene (lower), a known target of Rpb1. The input tracks are from the CUT&RUN input sample.
CUT and RUN Image 3: Digitonin Solution
CUT&RUN and ChIP assays were performed with HCT 116 cells and CTCF (D31H2) XP® Rabbit mAb #3418. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. Panel A compares enrichment of CTCF across chromosome 8 (upper), while Panel B compares enrichment at the MYC gene (lower), a known target of CTCF. The input tracks are from the CUT&RUN input sample.
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Product Usage Information

For the CUT&RUN assay, we recommend adding 25 μl of Digitonin Solution per 1 ml buffer. Not all cell lines exhibit the same sensitivity to digitonin, so it may be necessary to adjust the final volume of digitonin in the buffer to get optimal permeabilization of your specific cell line. Cell permeabilization can be tested by combining an equal volume of cell suspension and 0.4% Trypan Blue Stain. Optimal cell permeabilization will result in staining of >90% of the cell population.

Storage

Digitonin Solution is supplied in nuclease-free water. Store at -20°C. This product is stable for at least 12 months.

Product Description

The Digitonin Solution 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 completely thawed at 90-100°C for 5 minutes before using. Please keep on ice during use and store at -20°C when finished for the day.

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 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 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.
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To Purchase # 16359
Cat. # Size Qty. Price
16359L		 2.4 ml  (2 x 1.2 ml)
$ 112

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