H3K4me3 Antibody, SNAP-Certified™ for CUT&RUN and CUT&Tag

Product Description

This H3K4me3 (histone H3 trimethylated at lysine 4) antibody meets EpiCypher’s lot-specific SNAP-Certified™ criteria for specificity and efficient target enrichment in both  CUT&RUN and  CUT&Tag applications. This requires <20% cross-reactivity to related histone PTMs determined using the SNAP-CUTANA™ K-AcylStat Panel of spike-in controls . High target efficiency is confirmed by consistent genomic enrichment at varying cell inputs: 500k and 50k cells in CUT&RUN; 100k and 10k cells in CUT&Tag . High efficiency antibodies display similar peak structures and highly conserved genome-wide signal even at reduced cell numbers. This antibody targets histone H3K4me3, which is enriched at active promoters near transcription start sites (TSS) and promotes gene activation.

Validation Data - CUT & Tag

Figure 1: SNAP specificity analysis in CUT&Tag. CUT&Tag was performed and CUT&Tag sequencing reads were aligned to the unique DNA barcodes corresponding to each nucleosome in the K-MetStat panel (x-axis). Data are expressed as a percent relative to on-target recovery (H3K4me3 set to 100%). The antibody showed highly specific recovery of H3K4me3 spike-in nucleosomes at both 100k and 10k nuclei.

Figure 2: CUT&Tag genome-wide enrichment. CUT&Tag was performed and sequence reads were aligned to 18,793 annotated transcription start sites (TSSs ± 2 kbp). Signal enrichment was sorted from highest to lowest (top to bottom) relative to the H3K4me3 - 100k nuclei reaction (all gene rows aligned). High, medium, and low intensity are shown in red, yellow, and blue, respectively. H3K4me3 antibodies produced the expected enrichment pattern, which was consistent between 100k and 10k nuclei and greater than the IgG negative control.

Figure 3: H3K4me3 CUT&Tag representative browser tracks. CUT&Tag was performed and gene browser shots were generated using the Integrative Genomics Viewer (IGV, Broad Institute). H3K4me3 antibody tracks display sharp peaks at gene promoters, consistent with the biological function of this PTM. Similar results in peak structure and location were observed for both 100k and 10k nuclei inputs.

Validation Data - CUT & RUN

Figure 4: SNAP specificity analysis in CUT&RUN. CUT&RUN was performed and CUT&RUN sequencing reads were aligned to the unique DNA barcodes corresponding to each nucleosome in the K-MetStat panel (x-axis). Data are expressed as a percent relative to on-target recovery (H3K4me3 set to 100%). The antibody showed highly specific recovery of H3K4me3 spike-in nucleosomes at both 500k and 50k cells.

Figure 5: CUT&RUN genome-wide enrichment. CUT&RUN was performed and sequence reads were aligned to 18,793 annotated transcription start sites (TSSs ± 2 kbp). Signal enrichment was sorted from highest to lowest (top to bottom) relative to the H3K4me3 - 500k cells reaction (all gene rows aligned). High, medium, and low intensity are shown in red, yellow, and blue, respectively. H3K4me3 antibodies produced the expected enrichment pattern, which was consistent between 500k and 50k cells and greater than the IgG negative control.

Figure 6: H3K4me3 CUT&RUN representative browser tracks. CUT&RUN was performed and gene browser shots were generated using the Integrative Genomics Viewer (IGV, Broad Institute). H3K4me3 antibody tracks display sharp peaks at gene promoters, consistent with the biological function of this PTM. Similar results in peak structure and location were observed for both 500k and 50k cell inputs.