Description
Storage
All components in this kit are stable for 6 months when stored at the recommended temperature.
Protocol
Available protocols: CUT&Tag
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). This 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 Monoclonal Antibody #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 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 . CUT&Tag works very well for analyzing histone modifications, in addition to mapping some transcription factor and cofactor binding.
Alternate Names
C&T; CnT; CUT & Tag; cut and tag; CUT and Tag; CUT&Tag; cutandtag; histone modification
