CTCF Antibody

Western Blot: CTCF Antibody [NB500-177] - Detection of human CTCF by western blot. Samples: Whole cell lysate (50 ug) from Jurkat, HEK293T, and HeLa cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-CTCF antibody NB500-177 used for WB at 0.02 ug/ml. Detection: Chemiluminescence with an exposure time of 10 seconds.

Immunohistochemistry-Paraffin: CTCF Antibody [NB500-177] - Section of human small cell lung cancer. Antibody: Affinity purified rabbit anti-CTCF antibody (NB500-177) used at 1:1000. Secondary: HRP-conjugated goat anti-rabbit IgG.

Chromatin Immunoprecipitation: CTCF Antibody [NB500-177] - 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 20 micrograms of rabbit anti-CTCF antibody NB500-177. 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.

Western Blot: CTCF Antibody [NB500-177] - Detection of mouse CTCF by western blot. Samples: Whole cell lysate (50 ug) from NIH 3T3 cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-CTCF antibody NB500-177 used for WB at 0.02 ug/ml. Detection: Chemiluminescence with an exposure time of 75 seconds.

Immunohistochemistry-Paraffin: CTCF Antibody [NB500-177] - Section of mouse renal cell carcinoma. Antibody: Affinity purified rabbit anti-CTCF antibody (NB500-177) used at 1:1000. Secondary: HRP-conjugated goat anti-rabbit IgG.

Immunoprecipitation: CTCF Antibody [NB500-177] - Detection of human CTCF by western blot of immunoprecipitates. Samples: Whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) from Jurkat cells prepared using NETN lysis buffer. Antibodies: Affinity purified rabbit anti-CTCF antibody NB500-177 (lot NB500-177-6) used for IP at 6 ug per reaction. CTCF was also immunoprecipitated by a previous lot of this antibody (lot NB500-177-5). For blotting immunoprecipitated CTCF, NB500-177 was used at 0.02 ug/ml. Detection: Chemiluminescence with an exposure time of 10 seconds.

• Reactivity: Hu, Mu, Rt
• Applications: WB, IHC, IP, ChIP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 0.2 mg/ml

CTCF Antibody Summary

• Immunogen: The immunogen recognized by this antibody maps to a region between residues 650 and 700 of human CCCTC-binding factor using the numbering given in entry NP_006556.1 (GeneID 10664).
• Predicted Species: Rat (100%). Backed by our 100% Guarantee.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: CTCF
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation (ChIP) 1:10-1:500
  • Immunohistochemistry 1:500 - 1:2000
  • Immunohistochemistry-Paraffin 1:500 - 1:2000
  • Immunoprecipitation 2-10 ug/mg lysate
  • Western Blot 1:2500-1:10000
• Application Notes: Epitope retrieval with Tris-EDTA pH9.0 is recommended for FFPE tissue sections. Chip, WB, IP reacivity reported in (PMID: 24741094).

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: TBS and 0.1% BSA
• Preservative: 0.09% Sodium Azide
• Concentration: 0.2 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for CTCF Antibody

• 11 zinc finger transcriptional repressor
• 11-zinc finger protein
• 11-Zinc Finger Protein2
• CCCTC-binding factor (zinc finger protein)
• CCCTC-binding Factor
• CTCF
• CTCFL paralog
• transcriptional repressor CTCF

Background

The vertebrate insulator protein, CCCTC binding factor (CTCF) is a highly conserved, ubiquitously expressed DNA-binding protein that has widespread capabilities in gene regulation. CTCF contains 11 zinc finger motifs and it is thought that CTCF uses different combinations of zinc fingers to recognise a variety of specific sequences on the DNA to control several important genes. CTCF plays dual roles in cell growth regulatory processes and epigenetic functions. CTCF is a negative regulator of cell proliferation and is located in a region of the chromosome that is frequently deleted in human cancers. The CTCF protein is altered in some breast and prostate tumours. Substitutions from one amino acid to another in the DNA-binding domain change the whole spectrum of the genes regulated by CTCF which can result in cancer development. CTCF was first identified as one of the nuclear factors binding to the regulatory regions of the c-myc gene. Further study of this protein revealed roles in transcriptional repression, insulator function, and imprinting genetic information. Among the multitude of normal CTCF functions, its involvement in partitioning the genome into active or inactive domains by means of a chromatin insulator function has received particular attention. As an insulator CTCF helps to define the boundaries between the neighbouring genes. Strikingly, all vertebrate chromatin insulators identified so far interact with CTCF.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for CTCF Antibody (NB500-177)(2)

Publications using NB500-177

• Rivero-Hinojosa S, Pugacheva EM, Kang S et al. The combined action of CTCF and its testis-specific paralog BORIS is essential for spermatogenesis Nature Communications 2021-06-22 [PMID: 34158481] (Chemotaxis, Immunoprecipitation, Western Blot, Human)
• Martinez FP, Cruz R, Lu F et al. CTCF Binding to the First Intron of the Major Immediate-Early (MIE) Gene of Human Cytomegalovirus (HCMV) Negatively Regulates MIE Gene Expression and HCMV Replication. J. Virol. 2014-04-16 [PMID: 24741094] (WB, Chemotaxis, IP, Human)

Bioinformatics

• Gene Symbol: CTCF
• Entrez:
  • 10664(Human)
• Uniprot:
  • P49711(Human)