Histone H3K4me2 Antibody, SNAP-Certified™ for CUT&RUN

EpiCypher是一家为表观遗传学和染色质生物学研究提供高质量试剂和工具的专业制造商。EpiCypher推出的CUT&RUN级别的Histone H3K4me2 Antibody符合EpiCypher的批次特异性SNAP-CertifiedTM标准，在CUT&RUN中具有特异性和高效的靶点富集。通过SNAP-CUTANA™K-MetStat Panel的spike-in对照确定(EpiCypher 19-1002，Fig. 1)与相关组蛋白PTM的交叉反应性<20%，在500k和50k起始细胞中一致的基因组富集证实了高靶向效率(Fig. 2-4)。该抗体靶向的H3K4me2在转录活性基因的启动子和细胞发育过程中表达的基因中富集[1]。

[1] Pekowkska et al. Genome Res. (2010). PMID: 20841431.

产品详情

反应种属: Human, Mouse, Wide Range (Predicted)

宿主来源: Rabbit

实验应用: CUT&RUN, ICC, WB

免疫原: A synthetic peptide corresponding to histone H3 dimethylated at lysine 4

克隆性: Monoclonal

表达系统: HEK293 cells

保存温度: Stable for 1 year at -20°C from date of receipt

运输温度: Frozen cold packs.

产品形式: Protein A affinity-purified antibody in PBS, 0.09% sodium azide, 1% BSA, and 50% glycerol

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验证数据

Figure 1: SNAP specificity analysis in CUT&RUN

CUT&RUN was performed as described above. 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 (H3K4me2 set to 100%).

Figure 2: CUT&RUN genome wide enrichment

CUT&RUN was performed as described above. 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 H3K4me2 - 50k cells sample (all gene rows aligned). High, medium, and low intensity are shown in red, yellow, and blue, respectively. H3K4me2 antibody produced the expected TSS enrichment pattern, which was consistent between 500k and 50k cells and greater than the IgG negative control.

Figure 3: H3K4me2 CUT&RUN representative browser tracks

CUT&RUN was performed as described above. Gene browser shots were generated using the Integrative Genomics Viewer (IGV, Broad Institute). Similar results in peak structure and location were observed for both cell inputs.

Figure 4: Antibody efficiency analysis in CUT&RUN using cell input correlation

CUT&RUN was performed as described above. Genome-wide correlation analysis was performed to compare H3K4me2 antibody enrichment 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 H3K4me2 antibody are highly correlated between the two cell inputs (Pearson correlation r = 0.942), indicating high efficiency of H3K4me2 antibody target recovery.

Figure 5: Immunocytochemistry

ICC of HeLa cells using 2 µg/mL of H3K4me2 antibody (red). Actin filaments were labeled with fluorescein phalloidin (green).

Figure 6: Western blot data

Recombinant histone H3.3 (Lane 1) and acid extracts of HeLa cells (Lane 2) were blotted onto PVDF and probed with 0.025 µg/mL of H3K4me2 antibody.

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