Overview

  • Product nameAnti-CTCF antibody - ChIP Grade
    See all CTCF primary antibodies
  • Description
    Rabbit polyclonal to CTCF - ChIP Grade
  • Tested applicationsSuitable for: WB, IP, ChIP, IHC-P, EMSAmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Cow, Pig, Chimpanzee, Rhesus monkey, Gorilla, Orangutan, Elephant
  • Immunogen

    Synthetic peptide corresponding to a region between residues 650 and 700 of human CTCF (Peptide available as ab115042.)

  • Positive control
    • MCF7, 293T, NIH3T3 and HeLa whole cell lysates and Primary human lung fibroblasts.

Properties

Applications

Our Abpromise guarantee covers the use of ab70303 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/2500 - 1/10000. Detects a band of approximately 83 kDa (predicted molecular weight: 83 kDa).
IP Use at 2-5 µg/mg of lysate.
ChIP Use at an assay dependent concentration.
IHC-P 1/100 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
EMSA Use at an assay dependent concentration. PubMed: 22210889

Target

  • FunctionChromatin binding factor that binds to DNA sequence specific sites. Involved in transcriptional regulation by binding to chromatin insulators and preventing interaction between promoter and nearby enhancers and silencers. Acts as transcriptional repressor binding to promoters of vertebrate MYC gene and BAG1 gene. Also binds to the PLK and PIM1 promoters. Acts as a transcriptional activator of APP. Regulates APOA1/C3/A4/A5 gene cluster and controls MHC class II gene expression. Plays an essential role in oocyte and preimplantation embryo development by activating or repressing transcription. Seems to act as tumor suppressor. Plays a critical role in the epigenetic regulation. Participates to the allele-specific gene expression at the imprinted IGF2/H19 gene locus. On the maternal allele, binding within the H19 imprinting control region (ICR) mediates maternally inherited higher-order chromatin conformation to restrict enhancer access to IGF2. Plays a critical role in gene silencing over considerable distances in the genome. Preferentially interacts with unmethylated DNA, preventing spreading of CpG methylation and maintaining methylation-free zones. Inversely, binding to target sites is prevented by CpG methylation. Plays a important role in chromatin remodeling. Can dimerize when it is bound to different DNA sequences, mediating long-range chromatin looping. Mediates interchromosomal association between IGF2/H19 and WSB1/NF1 and may direct distant DNA segments to a common transcription factory. Causes local loss of histone acetylation and gain of histone methylation in the beta-globin locus, without affecting transcription. When bound to chromatin, it provides an anchor point for nucleosomes positioning. Seems to be essential for homologous X-chromosome pairing. May participate with Tsix in establishing a regulatable epigenetic switch for X chromosome inactivation. May play a role in preventing the propagation of stable methylation at the escape genes from X- inactivation. Involved in sister chromatid cohesion. Associates with both centromeres and chromosomal arms during metaphase and required for cohesin localization to CTCF sites. Regulates asynchronous replication of IGF2/H19.
  • Tissue specificityUbiquitous. Absent in primary spermatocytes.
  • Sequence similaritiesBelongs to the CTCF zinc-finger protein family.
    Contains 11 C2H2-type zinc fingers.
  • DomainThe 11 zinc fingers are highly conserved among vertebrates, exhibiting almost identical amino acid sequences. Different subsets or combination of individual zinc fingers gives the ability to CTCF to recognize multiple DNA target sites.
  • Post-translational
    modifications
    Sumoylated on Lys-74 and Lys-689; sumoylation of CTCF contributes to the repressive function of CTCF on the MYC P2 promoter.
  • Cellular localizationNucleus > nucleoplasm. Chromosome. Chromosome > centromere. May translocate to the nucleolus upon cell differentiation. Associates with both centromeres and chromosomal arms during metaphase. Associates with the H19 ICR in mitotic chromosomes. May be preferentially excluded from heterochromatin during interphase.
  • Information by UniProt
  • Database links
  • Alternative names
    • 11 zinc finger protein antibody
    • 11 zinc finger transcriptional repressor antibody
    • 11-zinc finger protein antibody
    • CCCTC binding factor (zinc finger protein) antibody
    • CCCTC binding factor antibody
    • CCCTC-binding factor antibody
    • Ctcf antibody
    • CTCF_HUMAN antibody
    • CTCFL paralog antibody
    • MRD21 antibody
    • Transcriptional repressor CTCF antibody
    see all

Anti-CTCF antibody - ChIP Grade images



  • Predicted band size : 83 kDa

    Samples: Whole cell lysate (50 µg) from CT26.WT, 4T1, Renca, and TCMK-1 cells.  ab70303 used at a dilution of 0.2 µg/ml.

    Detection: Chemiluminescence with an exposure time of 30 seconds.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma (left) and mouse renal cell carcinoma (right) tissues labelling CTCF with ab70303 at 1/100 (2µg/ml). Detection: DAB.
  • All lanes : Anti-CTCF antibody - ChIP Grade (ab70303) at 0.03 µg/ml

    Lane 1 : MCF7 whole cell lysate at 50 µg
    Lane 2 : MCF7 whole cell lysate at 15 µg
    Lane 3 : MCF7 whole cell lysate at 5 µg
    Lane 4 : 293T whole cell lysate at 50 µg
    Lane 5 : NIH3T3 whole cell lysate at 50 µg
    Lane 6 : HeLa whole cell lysate at 50 µg


    Predicted band size : 83 kDa
    Observed band size : 83 kDa


    Exposure time : 30 seconds
  • Detection of Human CTCF by Western Blot of Immunprecipitate. Lane 1: MCF7 whole cell lysates immunoprecipitated using ab70302 at 3µg/mg lysate (1 mg/IP; 1/4 of IP loaded/lane). Lane 2: MCF7 whole cell lysates immunoprecipitated using ab70303 at 3µg/mg lysate (1 mg/IP; 1/4 of IP loaded/lane). Lane 3: MCF7 whole cell lysates immunoprecipitated using ab70304 at 3µg/mg lysate (1 mg/IP; 1/4 of IP loaded/lane). Lane 4: control IgG was used for immunoprecipitation. All lanes: ab70303 at 0.03µg/ml. Detection: Chemiluminescence with exposure times of 10 seconds.
  • Analysis of Human CTCF by Chromatin Immunoprecipitation (ChIP). Primary human fibroblasts from lung (5 x 107 cells) were cross-linked with formaldehyde, sonicated, and immunoprecipitated with 20µg ab70303. The resulting ChIP DNA was quantified using real-time PCR with primers against the IGF2-H19 ICR or a control region in the HoxA cluster.

References for Anti-CTCF antibody - ChIP Grade (ab70303)

This product has been referenced in:
  • Kolodziej S  et al. PADI4 acts as a coactivator of Tal1 by counteracting repressive histone arginine methylation. Nat Commun 5:3995 (2014). WB ; Human . Read more (PubMed: 24874575) »
  • Srivastava S  et al. Vertebrate GAGA factor associated insulator elements demarcate homeotic genes in the HOX clusters. Epigenetics Chromatin 6:8 (2013). ChIP ; Mouse . Read more (PubMed: 23607454) »

See all 10 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (U2OS)
Permeabilization Yes - Methanol
Specification U2OS
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted May 05 2015

I've compiled some information below regarding potential primer sets for the ChIP antibodies we talked about, where information was lacking (verified in ChIP by other researches), I included links to abReviews and references that used the antibody in C...

Read More

Thank you for contacting Abcam regarding ab70303.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement.

To check the sta...

Read More

Thanks for your email.

Upon further enquiry I have found that the ChIP data shown on our online datasheet was submitted by users using our Abreview online customer review system. The content of the reviews can be accessed at http://www.abcam....

Read More

Are these primer sequences available?

Thanks for your help!

Merci beaucoup pour votre réponse.

Je comprends votre soucis. Comme j'ai mentionné dans mon émail précédant, si vous souhaitez tester un autre de nos anticorps en ChIP (en ayant l'espoir qu'il soit plus &eac...

Read More

Merci pour votre émail ainsi que pour votre patience.

Effectivement, la légende n'est pas une erreur. Ce le laboratoire qui avait eu testé cet anticorps en ChIP avait utilisé 20ug pour le test et fait l'image montr...

Read More

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issuedtwo free of charge replacements:

One vial of ab18554 is being replaced with one vial of ab9574.

Read More

Thank you for contacting Abcam regarding ab70303.


Regarding the immunogen used to generate this antibody, it is not specifically phosphorylated and thus this antibody does not specifically target phosphorylated CTCF. Thus phosphatase in...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Human Cell lysate - nuclear (LNCaP)
Negative control ChIP'ed with non-specific IgG, and tested by real-time qPCR in CTCF positive control regions such as H19ICR, c-Myc insulator and DM1 region.
Specification LNCaP
Detection step Real-time PCR
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde, RT
Positive control LNCaP cell chromatin ChIP'ed with CTCF antibody, and tested by real-time qPCR in CTCF positive control regions such as H19ICR, c-Myc insulator region and DM1 regions.
Username

Dr. Biswajyoti Sahu

Verified customer

Submitted Apr 03 2012

1-10 of 14 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"