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 1. Products /
 2. CUT&Tag Kits & Reagents /
 3. CUT&Tag Assay Kit


CUT&TAG ASSAY KIT #77552

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Figure 1. CUT&Tag, CUT&RUN, and ChIP-seq assays were performed with NCCIT cells
and Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb #9733, using this CUT&Tag
Assay Kit, the CUT&RUN Assay Kit #86652, or the SimpleChIP® Plus Enzymatic
Chromatin IP Kit (Magnetic Beads) #9005. DNA libraries were prepared using
CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415 for
CUT&Tag samples and DNA Library Prep Kit for Illumina Systems (ChIP-seq,
CUT&RUN) #56795 for ChIP-seq and CUT&RUN samples. The upper panel compares
enrichment around HoxA genes, while the lower panel compares enrichment around
HoxD genes, both are known target genes of H3K27me3.
Show more






Figure 1. CUT&Tag, CUT&RUN, and ChIP-seq assays were performed with NCCIT cells
and Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb #9733, using this CUT&Tag
Assay Kit, the CUT&RUN Assay Kit #86652, or the SimpleChIP® Plus Enzymatic
Chromatin IP Kit (Magnetic Beads) #9005. DNA libraries were prepared using
CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415 for
CUT&Tag samples and DNA Library Prep Kit for Illumina Systems (ChIP-seq,
CUT&RUN) #56795 for ChIP-seq and CUT&RUN samples. The upper panel compares
enrichment around HoxA genes, while the lower panel compares enrichment around
HoxD genes, both are known target genes of H3K27me3.
Show more






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To Purchase # 77552

Cat. # Size Qty. Price 77552S 1 Kit
24 assays

€916.00

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CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415

1 Kit

CTCF (D31H2) XP® Rabbit mAb #3418

20 µl

Di-Methyl-Histone H3 (Lys9) (D85B4) XP® Rabbit mAb #4658

20 µl










PRODUCT INFORMATION


PRODUCT DESCRIPTION

The CUT&Tag Assay Kit is designed to conveniently provide reagents needed to
perform up to 24 reactions. The kit has been optimized to work in fresh or
lightly fixed cells for all types of DNA-binding proteins, including histones,
transcription factors, and cofactors. In addition, the kit has been optimized to
work for histones in fresh or lightly fixed tissues. For analysis of
transcription factors and cofactors in tissues, we recommend using the CUT&RUN
Assay Kit #86652. If possible, we recommend using 100,000 cells or 1 mg of
tissue per CUT&Tag reaction. If starting cell number is limited, this kit is
validated to work with as few as 5,000 to 10,000 cells per reaction for histone
modification targets and as few as 20,000 cells per reaction for transcription
factors and cofactors. This kit is compatible with both whole cells and nuclei
as starting material. We have not found that using nuclei generates a stronger
signal or a higher signal-to-noise reaction compared to whole cells. A complete
assay can be performed in as little as one day. The CUT&Tag Assay Kit also
provides important controls to ensure a successful CUT&Tag experiment, including
positive control Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb #9751 and
negative controls Normal Rabbit IgG #2729 and Normal Mouse IgG #68860. The
negative control Normal Rabbit IgG #2729 or Normal Mouse IgG #68860 is optional
for the experiments, depending on the requirements of the peak calling algorithm
used. This kit is compatible with downstream Next Generation sequencing (NG-seq)
analysis, but not qPCR.




STORAGE

All components in this kit are stable for 6 months when stored at the
recommended temperature.


SPECIFICITY / SENSITIVITY

The CUT&Tag Assay Kit can be utilized with any CUT&Tag-validated antibody to
detect endogenous levels of protein-DNA interactions and histone modifications
in mammalian cells (see Figures 1–3). One CUT&Tag reaction can use between 5,000
to 250,000 cells or 1 to 5 mg of tissue of starting material (see Figures 5-7).
The kit is compatible with multiple species of antibodies, including rabbit and
mouse (see Figure 4). The positive control Tri-Methyl-Histone H3 (Lys4) (C42D8)
Rabbit mAb #9751 detects multiple species of tri-methyl histone H3 Lys4 protein,
including human, mouse, rat, and monkey.


BACKGROUND

Similar to Cleavage Under Targets and Release Using Nuclease (CUT&RUN), Cleavage
Under Targets and Tagmentation (CUT&Tag) is a powerful technique used for
probing protein-DNA interactions within the natural chromatin context of the
cell (1-3). CUT&Tag has many of the same advantages as the CUT&RUN assay in that
it provides a rapid, robust, and true low cell number protocol for detection of
protein-DNA interactions in the cell. In addition, the CUT&Tag assay adds an in
situ adaptor DNA ligation step carried out by the pAG-Tn5 enzyme, in which an
adaptor DNA is ligated directly to antibody-targeted chromatin DNA fragments in
the cell. As a result, subsequent DNA library preparation is much faster and
easier than library preparation following the CUT&RUN assay, free from DNA end
repair, A-tailing, and adaptor ligation in vitro. CUT&Tag works very well for
analyzing histone modifications, in addition to mapping some transcription
factor and cofactor binding.

 1. Kaya-Okur, H.S. et al. (2019) Nat Commun 10, 1930.
 2. Kaya-Okur, H.S. et al. (2020) Nat Protoc 15, 3264-3283.
 3. Henikoff, S. et al. (2021) Bio Protoc 11, e4043.



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FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
CUT&Tag provided under a license from Active Motif, Inc. under U.S. Patent No.
10,689,643 and 9,938,524, foreign equivalents, and child patents deriving
therefrom. For purchaser's internal research use only. May not be used for
resale, services, or other commercial use.
U.S. Patent No. 11,733,248, foreign equivalents, and child patents deriving
therefrom.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving
therefrom.
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