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.
BRD4 antibody produces CUT&RUN peaks primarily flanking transcription start sites (TSSs, Figure 1). BRD4 peaks show a large degree of overlap with BRG1/SMARCA4 peaks (Figure 2), as has been reported in the literature (Conrad et al., 2017).
Figure 1: BRD4 enrichment at annotated TSSs in CUT&RUN. CUT&RUN was performed using 500,000 K562 cells with BRD4 antibody as well as control antibodies (IgG negative control, EpiCypher 13-0042; H3K4me3 positive control, EpiCypher 13-0041). Sequencing reads were aligned to annotated TSSs (+/- 2 kbp) of 18,793 genes. High, medium, and low signal is ranked by intensity (top to bottom) and reflected by red, yellow, and blue colors, respectively. All rows aligned relative to H3K4me3 antibody.
Figure 2: BRD4 CUT&RUN peak enrichment and functional overlap. The CUT&RUN data from Figure 1 was subjected to peak calling using MACS2. BRD4 peaks overlapped with BRG1/ SMARCA4 antibody CUT&RUN peaks (EpiCypher 13-2002, top), as has been demonstrated in the literature (Conrad et al., 2017). Three representative loci show BRD4 peaks in relation to control and BRG1 antibodies (bottom, Integrative Genomics Viewer).
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.