WB, IP, IF-IC, ChIP
H M R Mk
Endogenous
140
Rabbit
#P49711
10664
Product Information
Product Usage Information
For optimal ChIP results, use 20 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:50 |
| Immunofluorescence (Immunocytochemistry) | 1:100 - 1:400 |
| Chromatin IP | 1:25 |
Storage
Specificity / Sensitivity
Species Reactivity:
Human, Mouse, Rat, Monkey
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the carboxy terminus of the human CTCF protein. Antibodies are purified by protein A and peptide affinity chromatography.
Background
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-A1 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).
- Klenova, E.M. et al. (2002) Semin Cancer Biol 12, 399-414.
- Klenova, E.M. et al. (1993) Mol Cell Biol 13, 7612-24.
- Loukinov, D.I. et al. (2002) Proc Natl Acad Sci USA 99, 6806-11.
- Mukhopadhyay, R. et al. (2004) Genome Res 14, 1594-602.
- Hark, A.T. et al. (2000) Nature 405, 486-9.
- Ohta, T. et al. (1999) Am J Hum Genet 64, 397-413.
- Chao, W. et al. (2002) Science 295, 345-7.
- Yu, W. et al. (2004) Nat Genet 36, 1105-10.
- El-Kady, A. and Klenova, E. (2005) FEBS Lett 579, 1424-34.
- Filippova, G.N. et al. (1998) Genes Chromosomes Cancer 22, 26-36.
- Filippova, G.N. et al. (2002) Cancer Res 62, 48-52.
- Vatolin, S. et al. (2005) Cancer Res 65, 7751-62.
- Hong, J.A. et al. (2005) Cancer Res 65, 7763-74.
- D'Arcy, V. et al. (2008) Br J Cancer 98, 571-9.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
WB: Western Blotting IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry) ChIP: Chromatin IP
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
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