- 询价
- Cell Signaling Technology
- 9728
- USA
- 2025年08月30日
- W, IF-IC, ChIP
- H,M,R,Mk
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- 详细信息
- 文献和实验
- 技术资料
- 抗体英文名：
Di-Methyl-Histone H3 (Lys27) (D18C8) XP Rabbit mAb__®
- 抗原：
synthetic peptide corresponding to the amino terminus of histone H3 in which Lys27 is di-methylated
- 应用范围：
W, IF-IC, ChIP
- 级别：
详见MSDS文件
- 供应商：
CST
- 库存：
大量
- 适应物种：
H,M,R,Mk
- 保质期：
详见说明书
- 是否单克隆：
1
- 保存条件：
-20°c
- 规格：
40 ul (4 western blots)/100 ul (10 western blots)/carrier free & custom formulation / quantity
| 规格： | 产品价格： | ￥请询价 | |
|---|---|---|---|
| 规格： | 40 ul (4 western blots) | 产品价格： | ￥请询价 |
| 规格： | 100 ul (10 western blots) | 产品价格： | ￥请询价 |
| 规格： | carrier free & custom formulation / quantity | 产品价格： | ￥请询价 |
pathway more info application references datasheet PDF MSDS PDF protocols
Applications Key: W=Western Blotting IF-IC=Immunofluorescence (Immunocytochemistry) ChIP=Chromatin IP
Reactivity Key: H=Human M=Mouse R=Rat Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
| Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|
| W IF-IC ChIP | H M R Mk | Endogenous | 17 | Rabbit IgG |
| Protocols | |
|---|---|
| Specificity / Sensitivity | Di-Methyl-Histone H3 (Lys27) (D18C8) XP® Rabbit mAb detects endogenous levels of histone H3 when di-methylated on Lys27. The antibody does show some cross-reactivity with mono-methylated Lys27, but does not cross-react with non-methylated or tri-methylated Lys27. In addition, the antibody does not cross-react with mono-methylated, di-methylated or tri-methylated histone H3 Lys4, Lys9, Lys36 or histone H4 Lys20. |
| Source / Purification | Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of histone H3 in which Lys27 is di-methylated. Western Blotting
Western blot analysis of extracts from various cell lines using Di-Methyl-Histone H3 (Lys27) (D18C8) XP® Rabbit mAb. IF-IC
Confocal immumunofluorescent analysis of HeLa cells using Di-Methyl-Histone H3 (Lys27) (D18C8) XP® Rabbit mAb (green). Actin filaments have been labled with DY-554 phalloidin (red). Chromatin IP
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4x106 HeLa cells and either 10 μl of Di-Methyl-Histone H3 (Lys27) (D18C8) XP® Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR, using SimpleChIP® Human GAPDH Exon 1 Primers #5516, SimpleChIP® Human MyoD1 Exon 1 Primers #4490, and SimpleChIP® Human AFM Intron 1 Primers #5098. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one. ELISA-Peptide
Di-Methyl-Histone H3 (Lys27) (D18C8) XP® Rabbit mAb specificity was determined by peptide ELISA. The graph depicts the binding of the antibody to pre-coated di-methyl histone H3 (Lys27) peptide in the presence of increasing concentrations of various competitor peptides. As shown, only the di-methyl histone H3 (Lys27) peptide competed away binding of the antibody. |
| Background | The nucleosome, made up of four core histone proteins (H2A, H2B, H3, and H4), is the primary building block of chromatin. Originally thought to function as a static scaffold for DNA packaging, histones have now been shown to be dynamic proteins, undergoing multiple types of post-translational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (1). Histone methylation is a major determinant for the formation of active and inactive regions of the genome and is crucial for the proper programming of the genome during development (2,3). Arginine methylation of histones H3 (Arg2, 17, 26) and H4 (Arg3) promotes transcriptional activation and is mediated by a family of protein arginine methyltransferases (PRMTs), including the co-activators PRMT1 and CARM1 (PRMT4) (4). In contrast, a more diverse set of histone lysine methyltransferases has been identified, all but one of which contain a conserved catalytic SET domain originally identified in the Drosophila Su(var)3-9, Enhancer of zeste, and Trithorax proteins. Lysine methylation occurs primarily on histones H3 (Lys4, 9, 27, 36, 79) and H4 (Lys20) and has been implicated in both transcriptional activation and silencing (4). Methylation of these lysine residues coordinates the recruitment of chromatin modifying enzymes containing methyl-lysine binding modules such as chromodomains (HP1, PRC1), PHD fingers (BPTF, ING2), tudor domains (53BP1), and WD-40 domains (WDR5) (5-8). The discovery of histone demethylases such as PADI4, LSD1, JMJD1, JMJD2, and JHDM1 has shown that methylation is a reversible epigenetic marker (9).
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| Application References | Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know ! |
| Companion Products |
For Research Use Only. Not For Use In Diagnostic Procedures. |
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文献和实验实际上将作为阴性对照使用。CST 推荐使用 Histone H3 (D2B12) XP® Rabbit mAb (ChIP Formulated) #4620。该抗体可检测与基因组中所有 DNA 序列结合的所有组蛋白 H3 类型（H3、H3.3、CENP-A）。因此，不管检测的位点活性状态如何，该抗体都能为 ChIP 实验提供通用的阳性对照。 阴性对照 阴性对照抗体（如正常兔 IgG）不能识别特异抗原表位，因此可用于检测非特异性结合。例如，如果阴性对照样品中的产物量等于特异
法所替代。利用微球菌核酸酶的切割特性（在染色质核小体之间的 DNA 更容易被切开），可以快速温和地将染色质 DNA 切断，且最大限度地保持了细胞原有染色质结构以及目的蛋白与 DNA 的结合状态，使 ChIP 的准确性和结果可信度大大提高。CST 推出的 SimpleChIP® 酶解法试剂盒，含有标准 ChIP 流程所需的整套试剂、ChIP 级蛋白质 G 微珠、阳性对照抗体（Histone H3(D2B12) XP® Rabbit mAb (ChIP Formulated) #4620）、阴性对照抗体、DNA
Reagents 100-bp DNA size ladder (Promega) Agarose Antisera (affinity-purified) These should be raised against histone peptides with mono-, di-, or trimethylation or acetylation at a specific lysine/arginine residue
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