- 询价
- Cell Signaling Technology
- 4658
- USA
- 2025年07月14日
- W, IP, IF-IC, ChIP
- H,M,R,Mk,Dm,X,Z,B,Pg,Sc,Ce
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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 (Lys9) (D85B4) XP® Rabbit mAb
- 抗原：
synthetic peptide corresponding to the amino terminus of histone H3 in which Lys9 is di-methylated
- 应用范围：
W, IP, IF-IC, ChIP
- 供应商：
CST
- 保质期：
详见说明书
- 适应物种：
H,M,R,Mk,Dm,X,Z,B,Pg,Sc,Ce
- 库存：
大量
- 级别：
详见MSDS文件
- 是否单克隆：
单克隆
- 保存条件：
-20°c
pathway more info application references datasheet PDF MSDS PDF protocols
Applications Key: W=Western Blotting IP=Immunoprecipitation IF-IC=Immunofluorescence (Immunocytochemistry) ChIP=Chromatin IP
Reactivity Key: H=Human M=Mouse R=Rat Mk=Monkey Dm=D. melanogaster X=Xenopus Z=Zebrafish B=Bovine Pg=Pig Sc=S. cerevisiae Ce=C. elegans
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 IP IF-IC ChIP | H M R Mk (Dm) (X) (Z) (B) (Pg) (Sc) (Ce) | Endogenous | 17 | Rabbit IgG |
| Protocols | |
|---|---|
| Specificity / Sensitivity | Di-Methyl-Histone H3 (Lys9) (D85B4) XP™ Rabbit mAb detects endogenous levels of histone H3 only when di-methylated on Lys9. The antibody does not cross-react with non-methylated, mono-methylated or tri-methylated Histone H3 Lys9. In addition, the antibody does not cross-react with histone H3 di-methylated on Lys4, Lys27, or Lys36 and histone H4 di-methylated on Lys20. |
| Source / Purification | Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of histone H3 in which Lys9 is di-methylated. Western Blotting
Western blot analysis of extracts from various cell lines using Di-Methyl-Histone H3 (Lys9) (D85B4) XP™ Rabbit mAb. IF-IC
Confocal immunofluorescent analysis of HeLa cells using Di-Methyl-Histone H3 (Lys9) (D85B4) XP™ Rabbit mAb(green). Actin filaments have been labeled with DY-554 phalloidin (red). ELISA
Di-Methyl Histone H3 (Lys9) (D85B4) XP™ Rabbit mAb specificity was determined by peptide ELISA. The graph depicts the binding of the antibody to pre-coated di-methyl histone H3 (Lys9) peptide in the presence of increasing concentrations of various competitor peptides. As shown, only the di-methyl histone H3 (Lys9) peptide competed away binding of the antibody. Chromatin IP
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 20 μl of Di-Methyl-Histone H3 (Lys9) (D85B4) 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 RPL30 Exon 3 Primers #7014, SimpleChIP® Human AFM Intron 1 Primers #5098, and SimpleChIP® Human α Satellite Repeat Primers #4486. 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. |
| 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).
|
| 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h）即可。2. 修复大法——不仅仅是「煮一煮」微波炉修复：简单易行效果好，CST 推荐使用微波炉完成修复。合适的修复液：根据抗体说明书使用合适的修复液。用柠檬酸修复后，切片需浸泡在修复液中，自然冷却；而用 EDTA 修复后，切片可直接从修复缸中取出，直接进行下一步。注：使用不同的修复方式和不同生产商的抗体检测人肺癌组织中 EGFR 的表达。第一排为 CST 的 EGF Receptor (D38B1) XP® Rabbit mAb（#4267），EDTA 的修复方式明显优于柠檬酸盐及胃蛋白
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
的染色强度。获得合适的染色强度后，将载玻片置于蒸馏水 (dH2O) 中以阻止进一步显色。抗体：Phospho–Stat3(Tyr705)(D3A7) XP® Rabbit mAb #9145样本：石蜡包埋的人类乳腺肿瘤显色检测：利用 SignalStain® DAB Substrate Kit #8059（左），竞争者提供的 DAB（中和右）CST 的 IHC/IF 明星产品推荐：生化试剂：Catalog NumberProduct NameProduct
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