H3K27me3 Antibody, SNAP-Certified™ for CUT&RUN and CUT&Tag
86 in stock
Powered by BiozThis H3K27me3 (histone H3 lysine 27 trimethyl) 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-MetStat Panel of spike-in controls (EpiCypher 19-1002, Figures 1 and 4). High target efficiency is confirmed by consistent genomic enrichment at varying cell inputs: 500k and 50k cells in CUT&RUN (Figures 2-3); 100k and 10k cells in CUT&Tag (Figures 5-6). High efficiency antibodies display similar peak structures at representative loci (Figures 2 and 5) and highly conserved genome-wide signal (Figures 3 and 6) even at reduced cell numbers. H3K27me3 is associated with repressed genes [Cai et al. 2021] and is anti-correlated with H3K4me3, a marker of active gene transcription enriched at transcription start sites (Figures 2 and 5).
Validation Data
Figure 1: SNAP specificity analysis in CUT&RUN
Figure 2: CUT&RUN genome-wide enrichment
Figure 3: H3K27me3 CUT&RUN representative browser tracks
CUT&RUN methods
CUT&RUN was performed on 500k and 50k K562 cells with the SNAP-CUTANA™ K-MetStat Panel (EpiCypher 19-1002) spiked-in prior to the addition of 0.5 µg of either IgG negative control (EpiCypher 13-0042), H3K4me3 positive control (EpiCypher 13-0041), or H3K27me3 antibodies. The experiment was performed using the CUTANA™ ChIC/CUT&RUN Kit v3.0 (EpiCypher 14-1048). Library preparation was performed with 5 ng of CUT&RUN enriched DNA (or the total amount recovered if less than 5 ng) using the CUTANA™ CUT&RUN Library Prep Kit (EpiCypher 14-1001/14-1002). Both kit protocols were adapted for high throughput Tecan liquid handling. Libraries were run on an Illumina NextSeq2000 with paired-end sequencing (2×50 bp). Sample sequencing depth was 16.8 million reads (IgG 500k cell input), 14.4 million reads (H3K4me3 500k cell input), 16.4 million reads (H3K27me3 500k cell input) and 11.6 million reads (H3K27me3 50k cell input). Data were aligned to the hg19 genome using Bowtie2. Data were filtered to remove duplicates, multi-aligned reads, and ENCODE DAC Exclusion List regions.
Figure 4: SNAP specificity analysis in CUT&Tag
Figure 5: CUT&Tag genome-wide enrichment
Figure 6: H3K27me3 CUT&Tag representative browser tracks
CUT&Tag methods
CUT&Tag was performed on 100k and 10k K562 nuclei with the SNAP-CUTANA™ K-MetStat Panel (EpiCypher 19-1002) spiked-in prior to the addition of 0.5 µg of either IgG negative control (EpiCypher 13-0042), H3K4me3 positive control (EpiCypher 13-0041), or H3K27me3 antibodies. The experiment was performed using the CUTANA™ Direct-to-PCR CUT&Tag Protocol. Libraries were run on an Illumina NextSeq2000 with paired-end sequencing (2×50 bp). Sample sequencing depth was 4.0 million reads (IgG 100k nuclei input), 3.7 million reads (H3K4me3 100k nuclei input), 6.9 million reads (H3K27me3 100k nuclei input) and 5.2 million reads (H3K27me3 10k nuclei input). Data were aligned to the hg19 genome using Bowtie2. Data were filtered to remove duplicates, multi-aligned reads, and ENCODE DAC Exclusion List regions.
- Type: Monoclonal [2084-1G5]
- Host: Rabbit
- Applications: CUT&RUN, CUT&Tag
- Reactivity: Human, Wide Range (Predicted)
- Format: Protein A affinity-purified
- Target Size: 15 kDa
- CUT&RUN: 0.5 µg per reaction
- CUT&Tag: 0.5 µg per reaction
- Uniprot ID H3.1 – P68431
- Alternate Names H3, H3/a, H3/b, H3/c, H3/d
