17-10044 ChIPAb+ CTCF - ChIP Validated Antibody and Primer Set

17-10044
25 assays  25 assays per set. Recommended use: ~2 μg of antibody per chromatin immunoprecipitation (dependent upon biological context).
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey Applications
      H WB, DB, IHC, IP, ELISA, Mplex, ChIP
      Description
      Catalogue Number17-10044
      Trade Name
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+ CTCF - ChIP Validated Antibody and Primer Set
      OverviewAll ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.
      The ChIPAb+ CTCF set includes the CTCF antibody, a negative control antibody (normal mouse IgG), and qPCR primers which amplify a 165 bp region of human H19 internal control region. The CTCF and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of CTCF-associated chromatin.
      Alternate Names
      • 11 zinc finger transcriptional repressor
      • 11-zinc finger protein
      • CCCTC-binding factor
      • CCCTC-binding factor (zinc finger protein)
      • CTCFL paralog
      • transcriptional repressor CTCF
      Background InformationCTCF, also known as CCCTC-binding factor, contains 11 zinc finger (ZF) binding domains that allow it to bind different DNA target sequences and proteins. This multifunctional protein can act as a transcriptional activator or repressor. As a transcriptional repressor, CTCF, binds to promoters of vertebrate c-myc gene. It also binds to the PLK and PIM1 promoters. CTCF may prevent the access of transcriptional activators to enhancers. CTCF also acts as a transcriptional activator of APP. It is involved in different aspects of gene regulation including promoter activation or repression, hormone-responsive gene silencing, methylation-dependent chromatin insulation, and genomic imprinting. Mutations of CTCF in this gene have been associated with invasive breast cancers, prostate cancers, and Wilms tumors, suggesting a role for CTCF may as a tumor suppressor.
      References
      Product Information
      FormatPurified
      Control
      • Includes negative control mouse IgG antibody and primers specific for human H19 ICR.
      PresentationAnti-CTCF, (mouse monoclonal IgG1κ. One vial containing 50 μg of protein G purified mouse monoclonal IgG1κ in buffer containing 0.1 M Tris-Glycine (pH 7.4, 150 mM NaCl) with 0.05% sodium azide, before the addition of glycerol to 30%. Store at -20°C.

      Normal Mouse IgG. Two vials containing 25 µg purified IgG in 25 µL storage buffer containing 0.1% sodium azide. Store at -20°C.

      ChIP Primers, Human H19 ICR. One vial containing 75 μL of 5 μM of each primer specific for human H19 internal control region. Store at -20°C.
      FOR: CCC ATC TTG CT GACC TCAC
      REV: AGA CCT GGG ACG TTT CTG TG
      Quality LevelMQ100
      Applications
      ApplicationThe ChIPAb+ CTCF set includes the CTCF antibody, a negative control antibody (normal mouse IgG) & qPCR primers which amplify a 165 bp region of human H19 internal control region.
      Key Applications
      • Western Blotting
      • Dot Blot
      • Immunohistochemistry
      • Immunoprecipitation
      • Enzyme Immunoassay (ELISA)
      • Multiplexing
      • Chromatin Immunoprecipitation (ChIP)
      Application NotesChromatin Immunoprecipitation:
      Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP with 15 min fixation) were subjected to chromatin immunoprecipitation using 2 µg of either Normal Mouse IgG or 2 µg of Anti-CTCF and the Magna ChIP™ G Kit (Cat. # 17-611).
      Successful immunoprecipitation of CTCF associated DNA fragments was verified by qPCR using ChIP Primers, Human H19 ICR as a positive locus, and a representative published negative locus (Wendt, K.S., et al., 2008) (Please see figures). Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated.
      Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.

      Western Blot Analysis:
      Representative lot data.
      K562 cell lysate was resolved by electrophoresis, transferred to PVDF membranes and probed with Anti-CTCF, clone 1.2.1.5.4 (1 μg/mL).
      Proteins were visualized using a Goat Anti-Mouse conjugated to HRP and a chemiluminescence detection system (Please see figures).

      The following molecular weights have been reported in the literature: 180 kDa (CTCF-180), 130 kDa (CTCF-130), 82 kDa (CTCF-82), 70 kDa (CTCF-70). See Klenova, E.M., et al, (1997) Nucleic Acids Res. 25, 466–473 and Torrano,V., et al, (2006) Journal of Cell Science 119, 1746-1759) for additional details.
      Please see molecular weight section for additional information.
      Biological Information
      ImmunogenHis-tagged recombinant protein corresponding to human CTCF.
      Clone1.2.1.5.4
      HostMouse
      SpecificityRecognizes CTCF, Mr ~130 kDa.
      IsotypeIgG1κ
      Species Reactivity
      • Human
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryThis gene is a member of the BORIS + CTCF gene family and encodes a transcriptional regulator protein with 11 highly conserved zinc finger (ZF) domains. This nuclear protein is able to use different combinations of the ZF domains to bind different DNA target sequences and proteins. Depending upon the context of the site, the protein can bind a histone acetyltransferase (HAT)-containing complex and function as a transcriptional activator or bind a histone deacetylase (HDAC)-containing complex and function as a transcriptional repressor. If the protein is bound to a transcriptional insulator element, it can block communication between enhancers and upstream promoters, thereby regulating imprinted expression. Mutations in this gene have been associated with invasive breast cancers, prostate cancers, and Wilms' tumors. [provided by RefSeq].
      Gene Symbol
      • CTCF
      Purification MethodProtein G Purified
      UniProt Number
      UniProt SummaryFunction: Transcriptional repressor binding to promoters of vertebrate c-myc gene. Also binds to the PLK and PIM1 promoters. May prevent the access of transcriptional activators to enhancers. Also acts as a transcriptional activator of APP. Involved in different aspects of gene regulation including promoter activation or repression, hormone-responsive gene silencing, methylation- dependent chromatin insulation, and genomic imprinting. Seems to act as tumor suppressor.SIZE: 727 amino acids; 82785 Da

      Subcellular Location Nucleus.

      Tissue Specificity:Ubiquitous. Absent in primary spermatocytes.

      Similarity: Belongs to the CTCF zinc-finger protein family. & Contains 11 C2H2-type zinc fingers.
      Molecular Weight~ 130 kDa
      Note: Poly(ADP-ribosyl)ated isoform, C-terminal truncation, or sumoylation will cause variable migration of CTCF protein on SDS–PAGE, which may present CTCF with different molecular weight on Western Blot. Modifications are dependent on cell type and growing condition.
      The following molecular weights have been reported in the literature: 180 kDa (CTCF-180), 130 kDa (CTCF-130), 82 kDa (CTCF-82), 70 kDa (CTCF-70). See Klenova,E.M.,et al, (1997) Nucleic Acids Res. 25, 466–473 and Torrano,V,et al, (2006) Journal of Cell Science 119, 1746-1759) for additional details.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceChromatin Immunoprecipitation:
      Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP with 15 min fixation) were subjected to chromatin immunoprecipitation using 2 µg of either Normal Mouse IgG or 2 µg of Anti-CTCF and the Magna ChIP™ G Kit (Cat. # 17-611). Successful immunoprecipitation of CTCF associated DNA fragments was verified by qPCR using ChIP Primers Human H19 ICR (Please see figures).
      Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsStable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
      Packaging Information
      Material Size25 assays
      Material Package25 assays per set. Recommended use: ~2 μg of antibody per chromatin immunoprecipitation (dependent upon biological context).
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      ChIPAb+ CTCF - ChIP Validated Antibody and Primer Set SDS

      Title

      Safety Data Sheet (SDS) 

      ChIPAb+ CTCF - ChIP Validated Antibody and Primer Set Certificates of Analysis

      TitleLot Number
      ChIPAb+ CTCF - 21085812108581
      ChIPAb+ CTCF - 23773892377389
      ChIPAb+ CTCF - 24335632433563
      ChIPAb+ CTCF - 20069372006937
      ChIPAb+ CTCF - 22468382246838
      ChIPAb+ CTCF - 31506493150649
      ChIPAb+ CTCF - 33804543380454
      ChIPAb+ CTCF - NG1800900NG1800900
      ChIPAb+ CTCF - NG1850728NG1850728
      ChIPAb+ CTCF - NG1912450NG1912450

      Brochure

      Title
      Advance your Epigenetics Research (MilliporeSigma)