Recombinant
RabMAb

Recombinant Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

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Overview

  • Product name

    Anti-CTCF antibody [EPR18253] - ChIP Grade
    See all CTCF primary antibodies

  • Description

    Rabbit monoclonal [EPR18253] to CTCF - ChIP Grade

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, ChIP, ICC/IF, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Positive control

    • WB: HeLa, LLC, RAW 264.7, HEK-293, HepG2, MCF7, C6, PC-12 and NIH/3T3 whole cell lysates; human fetal brain, fetal heart, fetal kidney and fetal spleen lysates; mouse brain, heart, kidney and spleen lysates; rat brain, heart, kidney and spleen lysates. IHC-P: Human endometrium, mouse liver and rat stomach tissues. ICC/IF: HeLa and NIH/3T3 cells. ChIP: Chromatin prepared from HeLa cells.

  • General notes

     

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

Our Abpromise guarantee covers the use of ab188408 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-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ChIP Use 2 µg for 25 µg of chromatin.
ICC/IF 1/2000.
WB 1/5000. Detects a band of approximately 140,130, 97, 80, 73, 70, 55 kDa (predicted molecular weight: 83, 46 kDa).

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

  • ChIP - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)
    ChIP - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

    Chromatin was prepared from HeLa (Human epithelial cells from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab188408 (blue), and 20µl of Anti Rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

  • Western blot - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)
    Western blot - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)
    All lanes : Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408) at 1/5000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 2 : LLC (Mouse lung carcinoma) whole cell lysate
    Lane 3 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 4 : HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate
    Lane 5 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysate
    Lane 6 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 83, 46 kDa


    Exposure time: 1 second


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Observed MW: 140, 97, 80, 73, 70, 55 kDa.

    The multiples bands observed in the WB have been reported in literature (PMID: 12878173, PMID: 8246978).

  • Western blot - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)
    Western blot - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)
    All lanes : Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408) at 1/1000 dilution

    Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 3 : NIH/3T3 (Mouse embryonic fibroblast cells) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 83, 46 kDa


    Exposure time: 1 second


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Observed MW: 140, 130, 97, 80, 73, 70, 55 kDa.

    The multiples bands observed in the WB have been reported in literature (PMID: 12878173, PMID: 8246978).

  • Western blot - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)
    Western blot - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)
    All lanes : Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408) at 1/1000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal heart lysate
    Lane 3 : Human fetal kidney lysate
    Lane 4 : Human fetal spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/100000 dilution

    Predicted band size: 83, 46 kDa


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Observed MW: 140, 130, 97, 80, 73, 70, 55 kDa.

    The multiples bands observed in the WB have been reported in literature (PMID: 12878173, PMID: 8246978).

  • Western blot - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)
    Western blot - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)
    All lanes : Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408) at 1/1000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Mouse kidney lysate
    Lane 4 : Mouse spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 83, 46 kDa


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Observed MW: 140, 130, 97, 80, 73, 70, 55 kDa.

    The multiples bands observed in the WB have been reported in literature (PMID: 12878173, PMID: 8246978).

  • Western blot - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)
    Western blot - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)
    All lanes : Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408) at 1/1000 dilution

    Lane 1 : Rat brain lysate
    Lane 2 : Rat heart lysate
    Lane 3 : Rat kidney lysate
    Lane 4 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 83, 46 kDa


    Exposure time: 2 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Observed MW 140, 130, 97, 80, 73, 70, 55 kDa.

    The multiples bands observed in the WB have been reported in literature (PMID: 12878173, PMID: 8246978).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

    Immunohistochemical analysis of paraffin-embedded human endometrium tissue labeling CTCF with ab188408 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on human endometrium is observed. Counter stained with Hematoxylin.

    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

    Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling CTCF with ab188408 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on hepatocytes of mouse liver is observed. Counter stained with Hematoxylin.

    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

    Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling CTCF with ab188408 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on the epithelium cells of rat stomach is observed. Counter stained with Hematoxylin.

    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)
    Immunocytochemistry/ Immunofluorescence - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling CTCF with ab188408 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).
    The negative controls are as follows:
    -ve control 1: ab188408 at 1/2000 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
    -ve control 2: ab7291 Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)
    Immunocytochemistry/ Immunofluorescence - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embyronic fibroblast cells) cells labeling CTCF with ab188408 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on NIH/3T3 cell line. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) seconday antibody at 1/1000 dilution (red).
    The negative controls are as follows:
    -ve control 1: ab188408 at 1/2000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
    -ve control 2:  Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

References

Publishing research using ab188408? Please let us know so that we can cite the reference in this datasheet.

ab188408 has not yet been referenced specifically in any publications.

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