Monoclonal antibody raised in rabbit against the region of histone H3 containing the trimethylated lysine 9 (H3K9me3), using a KLH-conjugated synthetic peptide.
| Lot | 001 |
|---|---|
| Concentration | 1 μg/μl |
| Species reactivity | Human, wide range expected. |
| Type | Monoclonal ChIP-seq grade |
| Purity | Affinity purified polyclonal antibody. |
| Host | Rabbit |
| Storage Conditions | Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles. |
| Storage Buffer | PBS containing 50% glycerol, 1% BSA and 0.09% azide. |
| Precautions | This product is for research use only. Not for use in diagnostic or therapeutic procedures. |
| Applications | Suggested dilution | References |
|---|---|---|
| ChIP/ChIP-seq * | 1 µg per IP | Fig 1, 2 |
| Western Blotting | 1:1,000 | Fig 3 |
*Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 0.5 - 5 µg per IP.
Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K9me3
ChIP was performed with the Diagenode antibody against H3K9me3 (cat. No. C15210014) on sheared chromatin from 500,000 HeLaS3 cells using the “iDeal ChIP-seq” kit (cat. No. C01010051). A titration of the antibody consisting of 0.5, 1, and 2 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. Quantitative PCR was performed with primers for the ZNF510 gene and the Sat2 satellite repeat, used as positive controls, and for the promoters of the GAPDH and EIF4A2 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
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Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K9me3
ChIP was performed on sheared chromatin from 500,000 HeLaS3 cells using 1 µg of the Diagenode antibody against H3K9me3 (cat. No. C15210014) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2A shows the signal distribution along the long arm of chromosome 19 and a zoomin to an enriched region containing several ZNF repeat genes. The arrows indicate two satellite repeat regions which exhibit a stronger signal. Figures 2B, C and D show the enrichment genomic regions surrounding the ZNF510 positive control target and at the KCNQ1and H19 imprinted genes, respectively.
Figure 3. Western blot analysis using the Diagenode monoclonal antibody directed against H3K9me3
Western blot was performed on whole cell extracts (40 µg) from HeLa cells using the Diagenode antibody against H3K9me3 (cat. No. C15210014). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is shown on the right, the marker (in kDa) is shown on the left.
 How to properly cite our product/service in your workWe strongly recommend using this: H3K9me3 monoclonal antibody (Hologic Diagenode Cat# C15210014 Lot# 001). Click here to copy to clipboard. Using our products or services in your publication? Let us know! |
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