Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

CTCF (D1A7) XP® Rabbit mAb #3417

imprinting   imprinting control region   insulator   methylation   sc-15914  

No. Size Price
3417S 100 µl ( 10 western blots ) ¥3,580.00 现货查询 购买询价
3417 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Rat,Monkey, Endogenous 140 Rabbit IgG
IP 1:50
IF-IC 1:100
ChIP 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP,


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

Specificity / Sensitivity

CTCF (D1A7) XP® Rabbit mAb detects endogenous levels of total CTCF protein. This antibody does not cross-react with BORIS.

CTCF (D1A7) XP® Rabbit mAb兔单抗能够检测内源性CTCF总蛋白水平。该抗体不与BORIS蛋白发生交叉反应。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the human CTCF protein.


Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 10 μl of CTCF (D1A7) 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 human c-Myc promoter primers, SimpleChIP® Human H19/Igf2 ICR Primers #5172, 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.

使用SimpleChIP®Enzymatic Chromatin IP Kit (Magnetic Beads) #9003,用4 x 106 HeLa细胞的交联染色质以及10 µl CTCF (D1A7) XP® Rabbit mAb或2 µl Normal Rabbit IgG #2729进行染色质免疫沉淀实验。使用human c-Myc promoter primers、SimpleChIP® Human H19/Igf2 ICR Primers #5172和SimpleChIP® Human α Satellite Repeat Primers #4486,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,这相当于一。

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using CTCF (D1A7) XP® Rabbit mAb.

使用CTCF (D1A7) XP® Rabbit mAb兔单抗,免疫印迹(Western blot)分析不同细胞中CTCF的蛋白水平。



Confocal immunofluorescent analysis of HeLa cells using CTCF (D1A7) XP® Rabbit mAb (green). Actin filaments are labeled with DY-554 phalloidin (red).

使用CTCF (D1A7) XP® Rabbit mAb 兔单抗(绿色)标记,共聚焦免疫荧光分析HeLa细胞。DY-554 phalloidin标记微丝蛋白(红色)。


CCCTC-binding factor (CTCF) and its paralog, the Brother of the Regulator of Imprinted Sites (BORIS), are highly conserved transcription factors that regulate transcriptional activation and repression, insulator function, and imprinting control regions (ICRs) (1-4). Although they have divergent amino and carboxy termini, both proteins contain 11 conserved zinc finger domains that work in combination to bind the same DNA elements (1). CTCF is ubiquitously expressed and contributes to transcriptional regulation of cell-growth regulated genes, including c-myc, p19/ARF, p16/INK4A, BRCA1, p53, p27, E2F1, and TERT (1). CTCF also binds to and is required for the enhancer-blocking activity of all known insulator elements and ICRs, including the H19/IgF2, Prader-Willi/Angelman syndrome, and Inactive X-Specific Transcript (XIST) anti-sense loci (5-7). CTCF DNA-binding is sensitive to DNA methylation, a mark that determines selection of the imprinted allele (maternal vs. paternal) (1). The various functions of CTCF are regulated by at least two different post-translational modifications. Poly(ADP-ribosyl)ation of CTCF is required for insulator function (8). Phosphorylation of Ser612 by protein kinase CK2 facilitates a switch of CTCF from a transcriptional repressor to an activator at the c-myc promoter (9). CTCF mutations or deletions have been found in many breast, prostate, and Wilms tumors (10,11). Expression of BORIS is restricted to spermatocytes and is mutually exclusive of CTCF (3). In cells expressing BORIS, promoters of X-linked cancer-testis antigens like MAGE-1A are demethylated and activated, but methylated and inactive in CTCF-expressing somatic cells (12). Like other testis specific proteins, BORIS is abnormally expressed in different cancers, such as breast cancer, and has a greater affinity than CTCF for DNA binding sites, detracting from CTCF’s potential tumor suppressing activity (1,3,13,14).

CCCTC-binding factor (CTCF)和它的横向同源物Brother of the Regulator of Imprinted Sites (BORIS)都是高度保守的转录因子,它们具有调节转录激活和抑制、绝缘子功能和印迹调控区域(Imprinting control regions,. ICRs) (1-4)。虽然它们有不同的氨基和羧基端,但是这两个蛋白都含有11个保守的锌指结构,这些结果可以联合结合到同一个DNA元件(1)。CTCF是广泛表达的,并且有助于细胞生长调节基因的转录调控,包括c-myc、p19/ARF、p16/INK4A、BRCA1、p53、p27、E2F1和TERT (1)。对于所有已知的绝缘子和ICRs包括H19/IgF2、Prader-Willi/Angelman syndrome和Inactive X-Specific Transcript (XIST) anti-sense loci的增强子的封闭活性,CTCF能够结合到它们并且是被需要的 (5-7)。CTCF与DNA的结合是对DNA的甲基化很敏感,甲基化是一个标记物,它能确定印记等位基因(母方vs.父方)的选择(1)。CTCF的不同功能通过至少两个不同的翻译后修饰来调节。CTCF的多聚腺苷酸二磷酸核糖基化反应对于绝缘子功能是需要的(8)。通过蛋白激酶CK2使在Ser612位点的磷酸化有助于在c-myc启动子中CTCF从一个转录抑制子到转录激活因子的开关作用(9)。CTCF的突变或截短在许多乳腺、前列腺和Wilms tumors中被发现(10,11)。BORIS的表达仅限于精母细胞,并且是与CTCF相互排斥的(3)。在表达BORIS的细胞中,X染色体相关的癌肿睾丸抗原例如MAGE-1A的启动子都被去甲基化并且激活,然而在CTCF表达的体细胞中是甲基化并且失活(12)。像其它睾丸特异性蛋白一样,BORIS在不同的癌症中不正常的表达例如乳腺癌,并且比CTCF对DNA结合位点有更大的亲和力,这可以减弱CTCF的潜在肿瘤抑制活性(1,3,13,14)。

  1. Klenova, E.M. et al. (2002) Semin Cancer Biol 12, 399-414.
  2. Klenova, E.M. et al. (1993) Mol Cell Biol 13, 7612-24.
  3. Loukinov, D.I. et al. (2002) Proc Natl Acad Sci USA 99, 6806-11.
  4. Mukhopadhyay, R. et al. (2004) Genome Res 14, 1594-602.
  5. Hark, A.T. et al. (2000) Nature 405, 486-9.
  6. Ohta, T. et al. (1999) Am J Hum Genet 64, 397-413.
  7. Chao, W. et al. (2002) Science 295, 345-7.
  8. Yu, W. et al. (2004) Nat Genet 36, 1105-10.
  9. El-Kady, A. and Klenova, E. (2005) FEBS Lett 579, 1424-34.
  10. Filippova, G.N. et al. (1998) Genes Chromosomes Cancer 22, 26-36.
  11. Filippova, G.N. et al. (2002) Cancer Res 62, 48-52.
  12. Vatolin, S. et al. (2005) Cancer Res 65, 7751-62.
  13. Hong, J.A. et al. (2005) Cancer Res 65, 7763-74.
  14. D'Arcy, V. et al. (2008) Br J Cancer 98, 571-9.

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