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H3K36me2 Antibody
Catalog Number
Format
Price
Other format
Monoclonal antibody raised in mouse against histone H3 dimethylated at lysine 36 (H3K36me2), using a KLH-conjugated synthetic peptide.
| Lot | 001-12 |
|---|---|
| Concentration | 1.0 µg/µl |
| Species reactivity | Human: positive. Other species: not tested. |
| Type | Monoclonal, ChIP-grade, CUT&Tag grade |
| Purity | Protein A purified monoclonal antibody. |
| Host | Mouse |
| Storage Conditions | Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles. |
| Storage Buffer | PBS containing 0.05% azide. |
| Precautions | This product is for research use only. Not for use in diagnostic or therapeutic procedures. |
| Applications | Suggested dilution | References |
|---|---|---|
| ChIP * | 1-2 µg/ChIP | Fig 1 |
| CUT&Tag | 1 µg | Fig 2 |
| ELISA | 1:3,000 | Fig 3 |
| Western Blotting | 1:1,000 - 1:2,000 |
* Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 µg per IP.
- Validation Data
Figure 1. ChIP results obtained with the monoclonal antibody directed against H3K36me2
ChIP assays were performed using human HeLa cells, a monoclonal antibody against H3K36me2 (cat. No. C15200182), and optimized PCR primer pairs for qPCR. ChIP was performed on sheared chromatin from 1 million cells. A titration consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for a genomic region upstream of the TGM2 gene, used as a positive control, and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
Figure 2. Cut&Tag results obtained with the monoclonal antibody directed against H3K36me2
Cut&Tag was performed on 50,000 K562 cells using 1 µg of the monoclonal antibody against H3K36me2 (cat. No. C15200182), the pA-Tn5 transposase (C01070001) and the iDeal Cut&Tag kit (cat. No. C01070021). The libraries were subsequently analyzed on an Illumina NovaSeq sequencer (2x50 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg38) using the BWA algorithm. Figure 2 shows the peak distribution in two genomic regions on chromosome 3 and X (figure 2A and 2B, respectively).
Figure 3. Cross reactivity of the monoclonal antibody directed against H3K36me2
To test the specificity, an ELISA was performed using a serial dilution of the monoclonal antibody against H3K36me2 (cat. No. C15200182). The wells were coated with peptides containing the unmodified H3K36 region as well as the mono-, di-, and trimethylated H3K36 and the trimethylated H3K9. Figure 3 shows a high specificity of the antibody for the peptide containing the modification of interest. - Publications
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Histone H3K36me2 and H3K36me3 form a chromatin platform essentialfor DNMT3A-dependent DNA methylation in mouse oocytes.
Yano Seiichi at al.
Establishment of the DNA methylation landscape of mammalian oocytes, mediated by the DNMT3A-DNMT3L complex, is crucial for reproduction and development. In mouse oocytes, high levels of DNA methylation occur exclusively in the transcriptionally active regions, with moderate to low levels of methylation in other regi...A histone H3.3K36M mutation in mice causes an imbalance of histonemodifications and defects in chondrocyte differentiation.
Abe, Shusaku and Nagatomo, Hiroaki and Sasaki, Hiroyuki and Ishiuchi,Takashi
Histone lysine-to-methionine (K-to-M) mutations have been identified as driver mutations in human cancers. Interestingly, these 'oncohistone' mutations inhibit the activity of histone methyltransferases. Therefore, they can potentially be used as versatile tools to investigate the roles of histone modifications. In ...A lncRNA regulates alternative splicing via establishment of a splicing-specific chromatin signature.
Gonzalez I, Munita R, Agirre E, Dittmer TA, Gysling K, Misteli T, Luco RF
Alternative pre-mRNA splicing is a highly cell type-specific process essential to generating protein diversity. However, the mechanisms responsible for the establishment and maintenance of heritable cell-specific alternative-splicing programs are poorly understood. Recent observations point to a role of histone modi...