Anti-CTCF antibody - ChIP Grade (ab70303)

Rabbit polyclonal CTCF antibody. Validated in WB, IP, IHC, EMSA, ChIP, CHIPseq and tested in Mouse, Human. Cited in 36 publication(s). Independently reviewed in 7 review(s).

Overview

  • Product name
    Anti-CTCF antibody - ChIP Grade
    See all CTCF primary antibodies
  • Description
    Rabbit polyclonal to CTCF - ChIP Grade
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, ChIP, IHC-P, CHIPseq, 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 lysate (ab150035) 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 100 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.
CHIPseq Use at an assay dependent concentration. PubMed: 28945252
EMSA Use at an assay dependent concentration. PubMed: 22210889

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
  • 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

  • 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.
  • 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

    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.

References

This product has been referenced in:
See all 36 Publications for this product

Customer reviews and Q&As

1-10 of 16 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted Mar 14 2017

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

Abcam user community

Verified customer

Submitted Feb 24 2017

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

Abcam user community

Verified customer

Submitted May 05 2015

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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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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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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Answer

Are these primer sequences available?

Thanks for your help!

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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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Answer

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 montré. Ils n'avaient pas testé des autres concentrations. Par contre nous avons des retours d'utilisateurs - les Abreviews - qui ont utilisé cet anticorps à soit 5 µg/µg chromatine soit à 1/200
https://www.abcam.com/index.html?datasheet=70303&tab=abreviews&intabreviewid=28894
https://www.abcam.com/index.html?datasheet=70303&tab=abreviews&intabreviewid=20853

Je suis désolée de ne pas pouvoir vous aider plus. Il semble que cet anticorps doit être utilisé assez concentré pour faire le ChIP. C'est pour l'instant le seul anticorps qui avait été testé en ChIP. Si par contrevous souhaitez tester un de nos autres anticorps contre le CTCF en ChIP, veuillez me contacter et nous pouvons regarder ce que nous pouvons faire ou conseiller.

J'espère que cette information vous est utile. Veuillez ne pas hésiter à nous recontacter pour des informations supplémentaires.

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Answer

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.

One vial of ab70303 is being replaced with one vial of ab126765.


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.

Read More

1-10 of 16 Abreviews or Q&A

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