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- 文献和实验
- 技术资料
- 抗体英文名:
Histone H3 (D2B12) XP Rabbit mAb (ChIP Formulated)__®
- 抗原:
synthetic peptide corresponding to the carboxy terminus of the human histone H3 protein
- 应用范围:
ChIP
- 适应物种:
H,M,R,Hm,Mk,C,Dm,X,Z,B
- 供应商:
CST
- 保质期:
详见说明书
- 级别:
详见MSDS文件
- 库存:
大量
- 是否单克隆:
1
- 保存条件:
-20°c
- 规格:
100 ul (10 immunoprecipitations)/carrier free & custom formulation / quantity
| 规格: | 产品价格: | ¥请询价 | |
|---|---|---|---|
| 规格: | 100 ul (10 immunoprecipitations) | 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: ChIP=Chromatin IP
Reactivity Key: H=Human M=Mouse R=Rat Hm=Hamster Mk=Monkey C=Chicken Dm=D. melanogaster X=Xenopus Z=Zebrafish B=Bovine
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 |
|---|---|---|---|---|
| ChIP | H M (R) (Hm) (Mk) (C) (Dm) (X) (Z) (B) | Endogenous | 17 | Rabbit IgG |
| Protocols |
|
|---|---|
| Specificity / Sensitivity | Histone H3 (D2B12) XP® Rabbit mAb (ChIP Formulated) detects endogenous levels of total histone H3 protein. This antibody does not cross-react with other histones. |
| Source / Purification | Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the carboxy terminus of the human histone H3 protein. Chromatin IP
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 10 μl of Histone H3 (D2B12) XP® Rabbit mAb (ChIP Formulated) 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 MYT-1 Exon 1 Primers #4493, 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 | Modulation of chromatin structure plays an important role in the regulation of transcription in eukaryotes. The nucleosome, made up of DNA wound around eight core histone proteins (two each of H2A, H2B, H3, and H4), is the primary building block of chromatin (1). The amino-terminal tails of core histones undergo various post-translational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (2-5). These modifications occur in response to various stimuli and have a direct effect on the accessibility of chromatin to transcription factors and, therefore, gene expression (6). In most species, histone H2B is primarily acetylated at Lys5, 12, 15, and 20 (4,7). Histone H3 is primarily acetylated at Lys9, 14, 18, 23, 27, and 56. Acetylation of H3 at Lys9 appears to have a dominant role in histone deposition and chromatin assembly in some organisms (2,3). Phosphorylation at Ser10, Ser28, and Thr11 of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis (8-10). Phosphorylation at Thr3 of histone H3 is highly conserved among many species and is catalyzed by the kinase haspin. Immunostaining with phospho-specific antibodies in mammalian cells reveals mitotic phosphorylation at Thr3 of H3 in prophase and its dephosphorylation during anaphase (11).
|
| 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
. However, to address in detail what happens at specific sites in vivo, chromatin immunoprecipitation (ChIP) is the method of choice. Here, we describe how ChIP can be performed on non-fixed chromatin from animal cells or tissues (fresh or frozen) to analyze histone
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