Histone

Histone H4K20me3 Antibody, SNAP-Certified™ for CUT&RUN

$475.00
SKU: 13-0054
Pack Size: 100 µg
  • Type:  Monoclonal
  • Target Size:  15 kDa
  • Format:  Affinity Purified IgG
  • Host:  Mouse
  • Reactivity:  Human, C. elegans
  • Applications:  CUT&RUN, WB

Description

This antibody meets EpiCypher’s lot-specific SNAP-Certified™ criteria for specificity and efficient target enrichment in CUT&RUN. This requires <20% cross-reactivity to related histone PTMs determined using the SNAP-CUTANA™ K-MetStat Panel of spike-in controls (EpiCypher 19-1002, Figure 1). High target efficiency is confirmed by consistent genomic enrichment at 500k and 50k starting cells (Figures 2-3).

Validation Data

Figure 1: SNAP specificity analysis in CUT&RUN
CUT&RUN was performed on 500k and 50k native K562 cells with the SNAP-CUTANA™ K-MetStat Panel (EpiCypher 19-1002) spiked-in prior to the addition of 0.5 µg H4K20me3 antibody. 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 (H4K20me3 set to 100%). Error bars represent mean ± SEM for the duplicate DNA barcodes in a single CUT&RUN experiment.

Figure 2: Antibody efficiency analysis in CUT&RUN using cell input correlation
Genome-wide correlation analysis was performed to compare H4K20me3 antibody enrichment in CUT&RUN using 500k cell and 50k cell inputs. The log of the number of reads per 75 bp binned region across the genome is plotted for both samples. CUT&RUN data generated using this H4K20me3 antibody are highly correlated between the two cell inputs (Pearson correlation r = 0.953), indicating high efficiency of H4K20me3 antibody target recovery.

Figure 3: H4K20me3 CUT&RUN representative tracks at 500k and 50k cells
Gene browser shots generated using the Integrative Genomics Viewer (IGV, Broad Institute) show representative loci for H4K20me3 antibody tested in CUT&RUN using 500k and 50k cells. Similar results in peak structure and location were observed for both cell inputs. Methods: CUT&RUN was performed using the CUTANA™ ChIC/CUT&RUN Kit v.2.0 (EpiCypher 14-1048) adapted to high throughout Tecan liquid handling. Library preparation was performed with 5 ng DNA using the NEBNext® Ultra™ II DNA Library Prep Kit for Illumina®. Libraries were run on an Illumina NextSeq2000 with paired-end sequencing (2x50bp). Sample sequencing depth was 4.6 million reads (IgG 50k cell input), 10.4 million reads (H4K20me3 50k cell input), and 5.7 million reads (H4K20me3 500k cell input).

Figure 4: Western blot data
Western analysis of H4K20me3 in HeLa cell lysates using H4K20me3 antibody at various dilutions.

Technical Information

Immunogen
A synthetic peptide corresponding to histone H4 trimethylated at lysine 20.
Formulation
Protein A affinity-purified antibody in PBS, with 0.1% sodium azide.
Storage and Stability
Stable for 1 year at -20°C from date of receipt.

Recommended Dilution

CUT&RUN: 0.5 µg per reaction

Western Blot (WB): 1:4,000

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