Does the CUT&Tag assay show a bias toward euchromatin or heterochromatin?

With the appropriate salt concentration in the tagmention buffer, we don't observe bias toward euchromatin or heterochromatin in our assays. In CUT&Tag, the target-specific antibody and the secondary antibody recruit the pAG-Tn5 and direct the tagmentation to the chromatin directly adjacent to the target-specific antibody, whether in euchromatin or heterochromatin. The active tethering of pAG-Tn5 to the chromatin allows for tagmentation to occur even in less accessible heterochromatin regions. If CST scientists observe non-specific tagmentation at open chromatin regions when they are validating an antibody for CUT&Tag, then the antibody is not approved for use in the CUT&Tag assay.

Our CUT&Tag Assay Kit #77552 has been shown to work well with antibodies against active, accessible euchromatic histone modifications (see Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb #9751) and activating transcription factors (see TCF4/TCF7L2 (C48H11) Rabbit mAb #2569), as well as repressive, inaccessible heterochromatic histone modifications (see Tri-Methyl-Histone H3 (Lys27) (C36B11) #9733) and polycomb repressor complex proteins that are associated with inactive heterochromatin (see Ezh2 (D2C9) XP® Rabbit mAb #5246).

Last updated: April 18, 2024