Rabbit Recombinant Multiclonal CTCF antibody. Carrier free. Suitable for WB, IHC-P, IHC-Fr, ICC/IF, Flow Cyt (Intra), IP, ChIP and reacts with Human, Mouse, Rat samples.
pH: 7.2 - 7.4
Constituents: 100% PBS
WB | IHC-P | IHC-Fr | ICC/IF | Flow Cyt (Intra) | IP | ChIP | |
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Human | Tested | Tested | Expected | Tested | Tested | Tested | Tested |
Mouse | Tested | Tested | Tested | Tested | Tested | Expected | Expected |
Rat | Tested | Tested | Tested | Expected | Expected | Expected | Expected |
Species | Dilution info | Notes |
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Species Human, Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Mouse | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Rat | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
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Species Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
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Species Human, Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Chromatin binding factor that binds to DNA sequence specific sites and regulates the 3D structure of chromatin (PubMed:18347100, PubMed:18654629, PubMed:19322193). Binds together strands of DNA, thus forming chromatin loops, and anchors DNA to cellular structures, such as the nuclear lamina (PubMed:18347100, PubMed:18654629, PubMed:19322193). Defines the boundaries between active and heterochromatic DNA via binding to chromatin insulators, thereby preventing interaction between promoter and nearby enhancers and silencers (PubMed:18347100, PubMed:18654629, PubMed:19322193). Plays a critical role in the epigenetic regulation (PubMed:16949368). Participates in the allele-specific gene expression at the imprinted IGF2/H19 gene locus (PubMed:16107875, PubMed:16815976, PubMed:17827499). 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 (By similarity). Mediates interchromosomal association between IGF2/H19 and WSB1/NF1 and may direct distant DNA segments to a common transcription factory (By similarity). Regulates asynchronous replication of IGF2/H19 (By similarity). Plays a critical role in gene silencing over considerable distances in the genome (By similarity). Preferentially interacts with unmethylated DNA, preventing spreading of CpG methylation and maintaining methylation-free zones (PubMed:18413740). Inversely, binding to target sites is prevented by CpG methylation (PubMed:18413740). Plays an important role in chromatin remodeling (PubMed:18413740). Can dimerize when it is bound to different DNA sequences, mediating long-range chromatin looping (PubMed:12191639). Causes local loss of histone acetylation and gain of histone methylation in the beta-globin locus, without affecting transcription (PubMed:12191639). When bound to chromatin, it provides an anchor point for nucleosomes positioning (PubMed:12191639). Seems to be essential for homologous X-chromosome pairing (By similarity). May participate with Tsix in establishing a regulatable epigenetic switch for X chromosome inactivation (PubMed:11743158). May play a role in preventing the propagation of stable methylation at the escape genes from X-inactivation (PubMed:11743158). Involved in sister chromatid cohesion (PubMed:12191639). Associates with both centromeres and chromosomal arms during metaphase and required for cohesin localization to CTCF sites (PubMed:18550811). Plays a role in the recruitment of CENPE to the pericentromeric/centromeric regions of the chromosome during mitosis (PubMed:26321640). Acts as a transcriptional repressor binding to promoters of vertebrate MYC gene and BAG1 gene (PubMed:18413740, PubMed:8649389, PubMed:9591631). Also binds to the PLK and PIM1 promoters (PubMed:12191639). Acts as a transcriptional activator of APP (PubMed:9407128). Regulates APOA1/C3/A4/A5 gene cluster and controls MHC class II gene expression (PubMed:18347100, PubMed:19322193). Plays an essential role in oocyte and preimplantation embryo development by activating or repressing transcription (By similarity). Seems to act as tumor suppressor (PubMed:12191639).
Transcriptional repressor CTCF, 11-zinc finger protein, CCCTC-binding factor, CTCFL paralog, CTCF
Rabbit Recombinant Multiclonal CTCF antibody. Carrier free. Suitable for WB, IHC-P, IHC-Fr, ICC/IF, Flow Cyt (Intra), IP, ChIP and reacts with Human, Mouse, Rat samples.
pH: 7.2 - 7.4
Constituents: 100% PBS
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
CTCF also known as the CCCTC-binding factor is a highly conserved zinc finger protein involved in transcriptional regulation and chromatin organization. It has a molecular weight of approximately 82 kDa. The CTCF protein plays a critical mechanical role in controlling the three-dimensional architecture of the genome by binding to specific DNA sequences and forming chromatin loops. It is widely expressed in various cell types across tissues where it acts as a transcriptional repressor and activator depending on the context. CTCF immunofluorescence techniques enable the visualization of its dynamic distribution and expression within the nucleus.
The multipurpose CTCF protein acts in coordinating the spatial organization of the genome. It functions as an insulator by regulating the boundaries between different chromosomal domains and controlling gene expression. CTCF operates within various complexes interacting with cohesin a vital protein complex that facilitates loop formation and influences genome architecture. This interaction helps in maintaining the integrity of the genome structure and proper chromatin insulation which are essential for normal gene function.
CTCF plays significant roles in epigenetic regulatory networks and transcriptional pathways. In the epigenetic landscape it influences gene expression through modulation of DNA methylation states at CpG islands interacting with proteins like DNA methyltransferases. In transcriptional pathways CTCF interacts with nuclear factor Y (NF-Y) which contributes to cell cycle regulation by modulating the expression of cell cycle genes. These pathways reflect CTCF's versatility in gene regulation and its influence on maintaining cellular homeostasis.
CTCF disruptions have been implicated in cancer and intellectual disabilities. Mutations or altered expression of CTCF can lead to tumorigenesis as CTCF acts as a tumor suppressor by controlling oncogene and tumor suppressor gene expression. In intellectual disabilities CTCF mutations affect brain development by disrupting the expression of neuronal genes. The protein's interaction with cohesin has links to disorders such as Cornelia de Lange syndrome where cohesin complex dysfunction parallels the phenotypes seen with CTCF aberrations.
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CTCF Western blot staining using rabbit Anti-CTCF antibody
This data was developed using Anti-CTCF antibody [RM1044] ab300639, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The multiples bands observed in the WB have been reported in literature (PMID: 12878173, PMID: 8246978).
All lanes: Western blot - Anti-CTCF antibody [RM1044] (Anti-CTCF antibody [RM1044] ab300639) at 1/1000 dilution
Lane 1: Human spleen lysate at 20 µg
Lane 2: Mouse heart lysate at 20 µg
Lane 3: Mouse spleen lysate at 20 µg
Lane 4: Mouse colon lysate at 20 µg
Lane 5: Rat heart lysate at 20 µg
Lane 6: Rat spleen lysate at 20 µg
Lane 7: Rat colon lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 83 kDa
Observed band size: 140 kDa, 130 kDa, 97 kDa, 80 kDa, 73 kDa, 55 kDa, 45 kDa
This data was developed using Anti-CTCF antibody [RM1044] ab300639, the same antibody clone in a different buffer formulation.
CTCF was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with Anti-CTCF antibody [RM1044] ab300639 at 1/30 dilution (2µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using Anti-CTCF antibody [RM1044] ab300639 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-CTCF antibody [RM1044] (Anti-CTCF antibody [RM1044] ab300639) at 1/30 dilution
All lanes: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Predicted band size: 83 kDa
Observed band size: 40 kDa, 130 kDa, 97 kDa, 80 kDa, 73 kDa, 55 kDa, 45 kDa
Exposure time: 10s
This data was developed using Anti-CTCF antibody [RM1044] ab300639, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human breast tissue labelling CTCF with Anti-CTCF antibody [RM1044] ab300639 at 1/10000 (0.053 µg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Nuclear staining on human breast is observed. The section was incubated with Anti-CTCF antibody [RM1044] ab300639 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: PBS was used instead of primary antibody followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).
This data was developed using Anti-CTCF antibody [RM1044] ab300639, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat colon tissue labelling CTCF with Anti-CTCF antibody [RM1044] ab300639 at 1/10000 (0.053 µg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Nuclear staining on rat colon. The section was incubated with Anti-CTCF antibody [RM1044] ab300639 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: PBS was used instead of primary antibody followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins was used.
This data was developed using Anti-CTCF antibody [RM1044] ab300639, the same antibody clone in a different buffer formulation.
Intracellular flow cytometric analysis of 4% paraformaldehyde fixed and 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling CTCF with Anti-CTCF antibody [RM1044] ab300639 at 1/500 dilution (0.1µg) (red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody.
This data was developed using Anti-CTCF antibody [RM1044] ab300639, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% paraformaldehyde fixed and 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) labeling CTCF with Anti-CTCF antibody [RM1044] ab300639 at 1/2000 dilution (0.266 μg/ml), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488, green) preadsorbed at 1:1000 (2 μg/mL). Confocal image showing nuclear staining in HeLa cell line.
Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (red) was used as a counterstain at 1:200 dilution (2.5 μg/ml). Nuclear counter satin is DAPI.
This data was developed using Anti-CTCF antibody [RM1044] ab300639, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse colon tissue labelling CTCF with Anti-CTCF antibody [RM1044] ab300639 at 1/10000 (0.053 µg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Nuclear staining on mouse colon. The section was incubated with Anti-CTCF antibody [RM1044] ab300639 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: PBS was used instead of primary antibody followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins was used.
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