Anti-CTCF antibody (ab70303)

Reviews (9)Q&A (9)References (46)

Key features and details

  • Rabbit polyclonal to CTCF
  • Suitable for: IHC-Fr, WB, IP, IHC-P
  • Reacts with: Mouse, Human
  • Isotype: IgG

Overview

  • Product name

    Anti-CTCF antibody
    See all CTCF primary antibodies

  • Description

    Rabbit polyclonal to CTCF

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-Fr, WB, IP, IHC-Pmore 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.)

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
IHC-Fr Use at an assay dependent concentration.
WB 1/2500 - 1/10000.

Detects a band of approximately 100 kDa (predicted molecular weight: 83 kDa).
IP Use at 2-5 µg/mg of lysate.
IHC-P 1/100 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function

    Chromatin 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 specificity

    Ubiquitous. Absent in primary spermatocytes.

  • Sequence similarities

    Belongs to the CTCF zinc-finger protein family.
    Contains 11 C2H2-type zinc fingers.

  • Domain

    The 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 localization

    Nucleus > 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

    see all

  • 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

Images

  • Western blot - Anti-CTCF antibody (ab70303)
    Western blot - Anti-CTCF antibody (ab70303)

    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) - Anti-CTCF antibody (ab70303)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTCF antibody (ab70303)
    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.
  • Western blot - Anti-CTCF antibody (ab70303)
    Western blot - Anti-CTCF antibody (ab70303)
    All lanes : Anti-CTCF antibody (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

    Exposure time: 30 seconds
  • Immunoprecipitation - Anti-CTCF antibody (ab70303)
    Immunoprecipitation - Anti-CTCF antibody (ab70303)
    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.

References (46)

Publishing research using ab70303? Please let us know so that we can cite the reference in this datasheet.

ab70303 has been referenced in 46 publications.

  • Rhodes JDP  et al. Cohesin Disrupts Polycomb-Dependent Chromosome Interactions in Embryonic Stem Cells. Cell Rep 30:820-835.e10 (2020). PubMed: 31968256
  • Oshima G  et al. DNA Methylation Controls Metastasis-Suppressive 14q32-Encoded miRNAs. Cancer Res 79:650-662 (2019). PubMed: 30538122
  • Jamshidi F  et al. Contribution of noncoding pathogenic variants to RPGRIP1-mediated inherited retinal degeneration. Genet Med 21:694-704 (2019). PubMed: 30072743
  • Hyle J  et al. Acute depletion of CTCF directly affects MYC regulation through loss of enhancer-promoter looping. Nucleic Acids Res 47:6699-6713 (2019). PubMed: 31127282
  • Kanehira K  et al. Fluorescence Enhancement Effect of TiO2 Nanoparticles and Application for Photodynamic Diagnosis. Int J Mol Sci 20:N/A (2019). PubMed: 31357730
View all Publications for this product

Customer reviews and Q&As

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1-10 of 18 Abreviews or Q&A

Western blot abreview for Anti-CTCF antibody

 Good
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (Heart)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
15 µg
Specification
Heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Abcam user community

Verified customer

Submitted Aug 19 2020

Immunohistochemistry (Frozen sections) abreview for Anti-CTCF antibody

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Heart)
Permeabilization
Yes - 0.2%Tween-20 in PBS
Specification
Heart
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
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Abcam user community

Verified customer

Submitted Aug 14 2020

Western blot abreview for Anti-CTCF antibody - ChIP Grade

 Good
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (DLD1)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
20 µg
Specification
DLD1
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Abcam user community

Verified customer

Submitted Mar 14 2017

ChIP abreview for Anti-CTCF antibody - ChIP Grade

 Excellent
Abreviews
abreview image
Application
ChIP
Sample
Human Cell lysate - nuclear (B Cells)
Negative control
IgG
Specification
B Cells
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 5 minute(s) and 0 second(s)
Specification of the cross-linking agent: Methanol free Formaldehyde
Positive control
Gene specific promoter
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Abcam user community

Verified customer

Submitted Feb 24 2017

Immunocytochemistry/ Immunofluorescence abreview for Anti-CTCF antibody - ChIP Grade

 Good
Abreviews
abreview image
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
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Abcam user community

Verified customer

Submitted May 05 2015

Question

Customer kindly contacted us regarding positve and negative primers for our ChIP grade antibodies.

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Answer

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 ChIP.

1. ab817: GAPDH (+), GAD1 (-), AFM (-), Factor VIII (-)

2. ab1791: GAPDH, RPL30, ALDOA, MYO-D, SERPINA, AFM, SAT2, SATa (each batch is verified against this primer set, an example image of one batch test is given on the datasheet)

3. ab7030: was verified in ChIP by other researchers submitting data through our abReview system and specific references. Their positive and negative controls used are given through the link below, as well as a "click to contact this user" link that allows you to contact the researchers if you would like more specific primer information. The specific references with ChIP listed in bold are good sources for positive and negative primers as well.
a) Abreview: (NFKBIA promoter (+), GAPDH (-)) https://www.abcam.com/HDAC3-antibody-ChIP-Grade-ab7030-reviews.html?intabreviewid=25057
b) Abreview: https://www.abcam.com/HDAC3-antibody-ChIP-Grade-ab7030-reviews.html?intabreviewid=18386
c) Specific references: https://www.abcam.com/HDAC3-antibody-ChIP-Grade-ab7030-references.html

4. ab111069 Untested in ChIP.

5. ab992: was verified in ChIP by other researchers (abReviews and specific references)
a) Specific references: https://www.abcam.com/Rad21-antibody-ChIP-Grade-ab992-references.html
b) Abreview: https://www.abcam.com/Rad21-antibody-ChIP-Grade-ab992-reviews.html?intabreviewid=14542
c) Abreview: (EBV 3+ (+), EBV1,2,4,5,6 (-), GAPDH (-), Cen (-)) https://www.abcam.com/Rad21-antibody-ChIP-Grade-ab992-reviews.html?intabreviewid=33942

Our ChIP primer sets:
https://www.abcam.com/index.html?pageconfig=resource&rid=10600

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Question

I received this product and it was hot when I opened it. I was advised to test it, but when I did it did not work.
Used in WB: tested various conditions for blocking and dilutions. Still no signal. I would like a replacement.

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Answer

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 status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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Question

Inquiry: Recently, we bought the antibody #70303 for ChIP assays. We are looking for suitable controls for the resulting ChIP DNA. Thus, please be so kind a send me the sequences of your primers (IGF2-H19 ICR and HoxA cluster).

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Answer

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 https://www.abcam.com/CTCF-antibody-ChIP-Grade-ab70303-reviews.html

The sequences of the primers were not provided by these reviewers. However, both reviewers have allowed other scientists to contact them via a link on the reviews. I recommend contacting the reviewers for those primer sequences.

Alternatively, there are alsoa few publications, whose citations and PubMed links are available on the 'Specific References' tab on the online datasheet, where ChIP is used within the paper. The authors may have provided primer sequences within these references.

I hope this is helpful. Please contact us again if you have any further questions.

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Question

We are looking for suitable controls for the resulting ChIP DNA. Thus, please be so kind a send me the sequences of your primers (IGF2-H19 ICR and HoxA cluster).

Read More
Answer

Are these primer sequences available?

Thanks for your help!

Read More

Question

Bonjour,

Merci beaucoup pour me donner la suite de la réponse.
Donc, ça fait des manipulations très chère, si c'est un tube entier par échantillon. Nous ne pouvons pas faire ça, c'est claire.
...

J'avais fait des testes et peut être je peux m'en sortir avec moins d'anticorps. J'avais vue les Abreviews avec 5 ul et 1/200, mais ça n'a pas suffit pour mes manipulations. Nous allons regarder ce qui est possible, avec notre budget.

Je vous remercie pour l'information.

Bonne soirée,

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Answer

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 économique pour le ChIP), je vous fais volontiers une offre de test comme décrite ici: https://www.abcam.com/collaborationdiscount

Je suis désolée de ne pas pouvoir vous aider autrement. Veuillez ne pas hésiter à nous recontacter, si vous avez des autres questions.

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1-10 of 18 Abreviews or Q&A

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