CTCF (D31H2) XP® Rabbit mAb #3418
- WB
- IP
- IHC
- IF
- ChIP
- C&R
- C&T
Supporting Data
| REACTIVITY | H M R Mk |
| SENSITIVITY | Endogenous |
| MW (kDa) | 140 |
| Source/Isotype | Rabbit IgG |
Application Key:
- WB-Western Blotting
- IP-Immunoprecipitation
- IHC-Immunohistochemistry
- IF-Immunofluorescence
- ChIP-Chromatin Immunoprecipitation
- C&R-CUT & RUN
- C&T-CUT & Tag
Species Cross-Reactivity Key:
- H-Human
- M-Mouse
- R-Rat
- Mk-Monkey
Product Information
Product Usage Information
The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
The CUT&Tag dilution was determined using CUT&Tag Assay Kit #77552.
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:50 |
| Immunohistochemistry (Paraffin) | 1:640 - 1:2560 |
| Immunofluorescence (Frozen) | 1:200 - 1:800 |
| Immunofluorescence (Immunocytochemistry) | 1:400 - 1:1600 |
| Chromatin IP | 1:50 |
| Chromatin IP-seq | 1:50 |
| CUT&RUN | 1:50 |
| CUT&Tag | 1:50 |
Storage
For a carrier free (BSA and azide free) version of this product see product #48859.
Protocol
Specificity / Sensitivity
CTCF (D31H2) XP® Rabbit mAb detects endogenous levels of total CTCF protein. This antibody does not cross-react with BORIS. This antibody nonspecifically labels the lamina propria of small intestine in fixed frozen mouse tissue by immunofluorescence.
Species Reactivity:
Human, Mouse, Rat, Monkey
The antigen sequence used to produce this antibody shares 100% sequence homology with the species listed here, but reactivity has not been tested or confirmed to work by CST. Use of this product with these species is not covered under our Product Performance Guarantee.
Species predicted to react based on 100% sequence homology:
Bovine, Horse
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the carboxy terminus of the human CTCF protein.
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.
Limited Uses
Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.
Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.