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

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: ChIP-sequencing, IHC-P, ChIP, ICC/IF, WB, ChIC/CUT&RUN-seqmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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. ChIP-seq: Chromatin prepared from Hela and Mouse Embryonic Fibroblasts cells. ChIC/CUT&RUN-Seq: HeLa and F9 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.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine)
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18253
Isotype

IgG
Research areas

Alternative Versions
- Anti-CTCF antibody [EPR18253] - ChIP Grade – BSA and Azide free (ab241391)
ChIP Related Products
- ChIP Kit (ab500)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human CTCF protein (ab153114)
Related Products
- ChIP Kit (Transcription factors, ChIP-seq) (ab270813)

The Abpromise guarantee

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
ChIP-sequencing		Use 4-8µg for 10⁷ cells.
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).
ChIC/CUT&RUN-seq		Use at an assay dependent concentration. 5 µg

Notes
ChIP-sequencing Use 4-8µg for 10⁷ cells.
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).
ChIC/CUT&RUN-seq Use at an assay dependent concentration. 5 µg

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.
Target information above from: UniProt accession P49711 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 10664 Human
- Entrez Gene: 13018 Mouse
- Entrez Gene: 83726 Rat
- Omim: 604167 Human
- SwissProt: P49711 Human
- SwissProt: Q61164 Mouse
- SwissProt: Q9R1D1 Rat
- Unigene: 368367 Human
- Unigene: 269474 Mouse
- Unigene: 18612 Rat
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

ChIC/CUT&RUN sequencing - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/µL, 2.5 x 10^5 F9 (Mouse embryonic testicular cancer cell line) cells and 5µg of ab188408 [EPR18253]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

Additional screenshots of mapped reads can be downloaded here.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
ChIP-sequencing - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

Chromatin was prepared from Mouse Embryonic Fibroblast cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 cells and 8 µg of [Anti-CTCF antibody]( ab188408). ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

Additional screenshots of mapped reads can be downloaded here.
ChIC/CUT&RUN sequencing - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/µL, 2 x 10^5 HeLa (Human cervix adenocarcinoma epithelial cell line) cells and 5 µg of ab188408 [EPR18253]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

The ChIP data was conducted on chromatin prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 HeLa cells and 8 µg of ab188408. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.

Additional screenshots of mapped reads can be downloaded here.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
ChIP-sequencing - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 30 µg of chromatin and 4 µg of ab188408. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. ChIP-Seq validation performed with ChIP-Kit Transcription Factors ChIP-Seq (ab270813).

Additional screenshots of mapped reads can be downloaded here.
ChIP-sequencing - Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408)

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 30 µg of chromatin and 4 µg of Anti-CTCF antibody [EPR18253] - ChIP Grade (ab188408). ChIP DNA was sequenced on the Illumina NextSeq 500 to a depth of 30 million reads. ChIP-Seq validation performed by Active Motif, Carlsbad, CA.

Additional screenshots of mapped reads can be downloaded here.
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)

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)

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)

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)

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)

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)

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)

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)

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)

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)

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

ChIP protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

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

ab188408 has been referenced in 8 publications.

Kayyar B et al. Regulation of Sox8 through lncRNA Mrhl-Mediated Chromatin Looping in Mouse Spermatogonia. Mol Cell Biol 42:e0047521 (2022). PubMed: 35412350
Xiao Y et al. Non-invasive diagnosis and surveillance of bladder cancer with driver and passenger DNA methylation in a prospective cohort study. Clin Transl Med 12:e1008 (2022). PubMed: 35968916
Azambuja AP & Simoes-Costa M The connectome of neural crest enhancers reveals regulatory features of signaling systems. Dev Cell 56:1268-1282.e6 (2021). PubMed: 33852891
Hou Y et al. CTCF Mediates Replicative Senescence Through POLD1. Front Cell Dev Biol 9:618586 (2021). PubMed: 33692996
Olbrich T et al. CTCF is a barrier for 2C-like reprogramming. Nat Commun 12:4856 (2021). PubMed: 34381034
Handa T et al. Chromatin integration labeling for mapping DNA-binding proteins and modifications with low input. Nat Protoc 15:3334-3360 (2020). PubMed: 32807906
Mei S et al. A functional variant in the boundary of a topological association domain is associated with pancreatic cancer risk. Mol Carcinog 58:1855-1862 (2019). PubMed: 31237042
Zhao X et al. Risk-Associated Long Noncoding RNA FOXD3-AS1 Inhibits Neuroblastoma Progression by Repressing PARP1-Mediated Activation of CTCF. Mol Ther 26:755-773 (2018). PubMed: 29398485

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

ChIP Related Products

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (8)

Customer reviews and Q&As

Post-translational
modifications