Monoclonal antibody raised in rabbit against the region of histone H3 containing the trimethylated lysine 36 (H3K36me3), 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 * | 0.5 - 1 µg per IP | Fig 1, 2 |
| Dot Blotting | 1:2,000 | Fig 3 |
| Western Blotting | 1:500 | Fig 4 |
*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 H3K36me3
ChIP was performed with the Diagenode antibody against H3K36me3 (cat. No. C15210013) 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 specific for the CCT5 coding region, used as positive control, and for a region upstream the ACTB promoter, used as negative control. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K36me3
ChIP was performed on sheared chromatin from 500,000 HeLaS3 cells using 1 µg of the Diagenode antibody against H3K36me3 (cat. No. C15210013) 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 2 shows the H3K36me3 signal along the complete sequence and 1.5 Mb region of human chromosome 1 (figure 2A and B), in a genomic region on chromosome X surrounding the EIF2S3 gene (figure 2C) and in a genomic region surrounding the CCT5 positive control (figure 2D).
Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K36me3
To test the cross reactivity of the Diagenode antibody against H3K36me3 (cat. No. C15210013), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K36. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:2,000. Figure 2 shows a high specificity of the antibody for the modification of interest.
Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K36me3
Western blot was performed on recombinant histone H3 (1 µg) and on histone extracts from HeLa cells (40 µg) using the Diagenode antibody against H3K36me3 (cat. No. C15210013). The antibody was diluted 1:500 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: H3K36me3 monoclonal antibody (Hologic Diagenode Cat# C15210013 Lot# 001). Click here to copy to clipboard. Using our products or services in your publication? Let us know! |
