CUTANA Compatible Antibodies meet the criteria for performance in Cleavage Under Targets and Release Using Nuclease (CUT&RUN) and/or Cleavage Under Targets and Tagmentation (CUT&Tag) approaches to genomic mapping.
Estrogen Receptor Alpha C-terminal (ER alpha C-term) antibody shows CUT&RUN peaks in response to estradiol stimulation (Figure 1) that overlap with known estrogen response element (ERE) binding motifs (Figure 2). Overlap is further observed with peaks from an antibody to a different ER alpha epitope (N-term; 13-2011-EPC) and NCOA3/SRC3 (13-2013-EPC), which coactivates ER-mediated transcription (1) (Figure 2).
Figure 1: ER alpha C-term enrichment in CUT&RUN. Serum-starved MCF7 cells were treated with 100 nM estradiol (E2) or vehicle control for 45 minutes. CUT&RUN was performed using 500,000 cells with 0.5 µg of each indicated antibody (gray text). Heatmap shows CUT&RUN enrichment in aligned rows ranked by intensity (top to bottom; relative to ER alpha C-term, 13-2012-EPC). Red indicates high localized enrichment and blue denotes background signal.
Figure 2: ER alpha C-term peak analysis in CUT&RUN. Peaks from the E2-treated samples in Figure 1 were called using MACS2. (A) The number of ER alpha C-term peaks which fall into distinct classes of functionally annotated genomic regions is plotted. (B) Homer analysis determined that the ERE consensus motif, represented as a sequence logo position weight matrix, was enriched under ER alpha C-term peaks. (C) The number of ER alpha C-term peaks containing consensus motifs from panel B is shown by Venn Diagram. (D) The number of ER alpha C-term peaks that overlap with ER alpha (N-term; 13-2011-EPC) and NCOA3/SRC3 (13-2013-EPC) antibodies are represented by Venn Diagram.
Every lot of a CUTANA Compatible antibody is tested in the indicated CUTANA approach using EpiCypher optimized protocols and determined to yield peaks that show a genomic distribution pattern consistent with reported function(s) of the target protein.