CUT&Tag pAG-Tn5 (Loaded) (#79561) Datasheet Without Images Cell Signaling Technology

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:

C&T

Product Information

Product Usage Information

Please refer to CUT&Tag Assay Kit #77552 for detailed use of this enzyme in the CUT&Tag assay. After cell permeabilization and primary and secondary antibody binding, resuspend cells in 50 μL of High Salt Digitonin Buffer containing 2 μL of pAG-Tn5 Enzyme (1:25 dilution). Incubate cell samples at room temperature for 1 hour, wash cells with High Salt Digitonin Buffer, and then perform the chromatin tagmentation.

Storage

Supplied in 22 mM HEPES pH 7.4, 44 mM NaCl, 44 μM EDTA, 0.4 mM DTT, 0.04% Triton X-100, and 50% glycerol. Store at –20°C and do not aliquot. This product is stable for 6 months.

Specificity / Sensitivity

Species Reactivity:

All Species Expected

Product Description

The CUT&Tag pAG-Tn5 (Loaded) enzyme provides enough enzyme to support 50 CUT&Tag assays. It is a fusion of Protein A and Protein G to Tn5, and is recombinantly produced in E. coli. This pAG-Tn5 has been loaded with the adaptor oligo that is compatible with NG-Sequencing for Illumina systems, so the genomic DNA fragments tagmented by CUT&Tag pAG-Tn5 (Loaded) are ready for PCR amplification and NG-seq. This enyzme is compatible with multiple species of antibodies, including both rabbit and mouse. This enzyme is validated using CUT&Tag Assay Kit #77552.

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.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Applications Key

C&T: CUT&Tag

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

Licensed from Active Motif, Inc. under certain pending and granted patents. For purchasers 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.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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