CTCF Polyclonal antibody

CTCF Polyclonal Antibody for WB, IHC, IF/ICC, IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

Human, mouse

Applications

WB, IHC, IF/ICC, IP, ELISA

Conjugate

Unconjugated

Cat no : 30428-1-AP

Synonyms

11 zinc finger protein, CCCTC binding factor, CTCF, CTCFL paralog, Transcriptional repressor CTCF

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Validation Data Gallery

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  WB analysis using 30428-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 30428-1-AP (CTCF antibody) at dilution of 1:8000 incubated at room temperature for 1.5 hours.

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  IP experiment of MCF-7 using 30428-1-AP

  IP result of anti-CTCF (IP:30428-1-AP, 4ug; Detection:30428-1-AP 1:8000) with MCF-7 cells lysate 1880 ug.

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  IHC staining of mouse lung using 30428-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse lung tissue slide using 30428-1-AP (CTCF antibody) at dilution of 1:800 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse lung using 30428-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse lung tissue slide using 30428-1-AP (CTCF antibody) at dilution of 1:800 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HepG2 using 30428-1-AP

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using CTCF antibody (30428-1-AP) at dilution of 1:1600 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).

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Tested Applications

• Positive WB detected in: HEK-293 cells, MCF-7 cells, RAW 264.7 cells
• Positive IP detected in: MCF-7 cells
• Positive IHC detected in: mouse lung tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:16000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:400-1:1600
• Immunofluorescence (IF)/ICC: IF/ICC : 1:800-1:3200

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

30428-1-AP targets CTCF in WB, IHC, IF/ICC, IP, ELISA applications and shows reactivity with Human, mouse samples.

• Tested Reactivity: Human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: CTCF fusion protein Ag32417
• Full Name: CCCTC-binding factor (zinc finger protein)
• Calculated Molecular Weight: 83 kDa
• Observed Molecular Weight: 110-150 kDa
• GenBank Accession Number: BC014267
• Gene Symbol: CTCF
• Gene ID (NCBI): 10664
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Transcriptional insulators are DNA elements that set boundaries on the actions of enhancer and silencer elements and thereby organize the eukaryotic genome into regulatory domains. All vertebrate insulators appear to use the versatile CTCF protein. CTCF uses various combinations of its 11 zinc fingers to recognize a variety of unrelated DNA sequences. Once bound to DNA, CTCF can function as a transcriptional insulator, repressor, or activator, depending on the context of the binding site [PMID:12787766,15454938]. In vertebrates, this 11 zinc-finger protein is shown to be crucial in processes of epigenetic imprinting, X chromosome inactivation , and associated with various complex human diseases including cancer and diabetes [PMID:23139640]. The calcualted molecular weight of CTCF is 83 kDa, but stimulation of human corneal epithelial cells with hypoxic stress suppressed a high molecular mass form of CTCF (150 kDa), but not a lower molecular weight form of CTCF (130 kDa)(PMID: 22354964), and there are multiple isoforms of CTCF with molecular masses of 55, 70, 73, 80, 97, and 130 kDa have been observed (PMID: 12878173).