Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

CHD1 (D8C2) Rabbit mAb #4351

No. Size Price
4351S 100 µl ( 10 western blots ) ¥3,100.00 现货查询 购买询价
4351 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 220 Rabbit IgG
IP 1:50
ChIP 1:100

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, ChIP=Chromatin IP,

Homology

Species predicted to react based on 100% sequence homology: Bovine, Dog, Pig,

Specificity / Sensitivity

CHD1 (D8C2) Rabbit mAb recognizes endogenous levels of total CHD1 protein.

CHD1 (D8C2) Rabbit mAb兔单抗能够检测内源性CHD1总蛋白水平。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human CHD1 protein.

通过合成的人源CHD1蛋白羧基端相应的多肽片段去免疫动物从而制备出此单克隆抗体。

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using CHD1 (D8C2) Rabbit mAb.

使用CHD1 (D8C2) Rabbit mAb兔单抗,免疫印迹(Western blot)分析不同细胞中CHD1 (D8C2)的蛋白水平。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 mES cells and either 5 μl of CHD1 (D8C2) 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® Mouse Oct-4 Promoter Primers #4653, SimpleChIP® Mouse RPL30 Intron 2 Primers #7015, and SimpleChIP® Mouse MYT-1 Promoter Primers #8985. 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.

使用SimpleChIP®Enzymatic Chromatin IP Kit (Magnetic Beads) #9003,用4 x 106 mES细胞的交联染色质以及5 µl CHD1 (D8C2) Rabbit mAb或2 µl Normal Rabbit IgG #2729进行染色质免疫沉淀实验。使用SimpleChIP® Mouse Oct-4 Promoter Primers #4653、SimpleChIP® Mouse RPL30 Intron 2 Primers #7015和SimpleChIP® Mouse MYT-1 Promoter Primers #8985,浓缩的DNA通过real-time PCR定量。在每个样品中免疫沉淀DNA的数量被当做一个相对于总input chromatin的数量的信号,相当于一个。

Background

Chromodomain-helicase-DNA-binding domain (CHD) proteins have been identified in a variety of organisms (1,2). This family of proteins, which consists of nine members, has been divided into three separate subfamilies: subfamily I (CHD1 and CHD2), subfamily II (CHD3 and CHD4), and subfamily III (CHD5, CHD6, CHD7, CHD8, and CHD9). All of the CHD proteins contain two tandem N-terminal chromodomains, a SWI/SNF-related ATPase domain, and a C-terminal DNA binding domain (1,2). The chromodomains facilitate binding to methylated lysine residues of histone proteins and confer interactions with specific regions of chromatin. The SWI/SNF-related ATPase domain utilizes the energy from ATP hydrolysis to modify chromatin structure. CHD1 is a euchromatic protein that associates with the promoters of active genes, and is required for the maintenance of open chromatin and pluripotency in embryonic stem cells (3-6). The two chromodomains of CHD1 facilitate its recruitment to active genes by binding to methyl-lysine 4 of histone H3, a mark associated with transcriptional activation (4-6). Yeast CHD1 is a component of the SAGA and SLIK histone acetyltransferase complexes, and is believed to link histone methylation with histone acetylation during transcriptional activation (6). The CHD2 protein is not well characterized; however, mouse knockout studies suggest important functions in development and tumor suppression. Homozygous CHD2 knockout mice exhibit delayed growth and perinatal lethality (7). Heterozygous knockout mice show increased mortality and gross organ abnormalities, in addition to increased extramedullary hematopoiesis and susceptibility to lymphomas (7,8). CHD2 mutant cells are defective in hematopoietic stem cell differentiation and exhibit aberrant DNA damage responses (8).

Chromodomain-helicase-DNA-binding domain (CHD)蛋白已经在多种生物中被鉴定(1,2)。由九个成员组成的蛋白家族已经被分为三个亚家族:subfamily I (CHD1和CHD2)、subfamily II (CHD3和CHD4)和subfamily III (CHD5、CHD6、CHD7、CHD8、CHD9)。所有CHD蛋白包含两个串联的N端chromodomains、一个SWI/SNF相关的ATPase结构域和一个C端DNA结合区域 (1,2)。chromodomains有助于结合到组蛋白的已甲基化赖氨酸残基,并且可与染色质的特异区域相互作用。SWI/SNF相关的ATPase区域可利用来自ATP水解的能量去修饰染色质结构。CHD1是一个常染色质蛋白,它与活化基因的启动子有关联,并且该蛋白对于在胚胎干细胞中染色质开放性和多能性的维持是需要的(3-6)。CHD1蛋白的两个chromodomains有助于它到活化基因的招募,该过程通过结合到histone H3的methyl-lysine 4位点,它是一个与转录激活相关联的标记物(4-6)。酵母CHD1蛋白是SAGA和SLIK组蛋白乙酰化转移酶复合物的一个成分,并且被认为在转录激活中联系着组蛋白甲基化和组蛋白乙酰化作用(6)。CHD2蛋白不具有明显特征;然而,小鼠基因敲除研究认为在发育和肿瘤抑制中有重要作用。纯合子CHD2敲除老鼠显示延迟生长和围产期致死(7)。杂合子敲除老鼠显示除了增加的髓外造血和易感性淋巴瘤之外,还出现死亡率增加和所有器官不正常(7,8)。CHD2突变细胞在造血干细胞分化中缺失,并且展示异常的DNA损伤反应(8)。

  1. Hall, J.A. and Georgel, P.T. (2007) Biochem Cell Biol 85, 463-76.
  2. Marfella, C.G. and Imbalzano, A.N. (2007) Mutat Res 618, 30-40.
  3. Gaspar-Maia, A. et al. (2009) Nature 460, 863-8.
  4. Sims, R.J. et al. (2005) J Biol Chem 280, 41789-92.
  5. Flanagan, J.F. et al. (2005) Nature 438, 1181-5.
  6. Pray-Grant, M.G. et al. (2005) Nature 433, 434-8.
  7. Marfella, C.G. et al. (2006) J Cell Physiol 209, 162-71.
  8. Nagarajan, P. et al. (2009) Oncogene 28, 1053-62.

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Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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