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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| H | WB, DB, IHC, IP, ELISA, Mplex, ChIP |
| Description | |
|---|---|
| Catalogue Number | 17-10044 |
| Trade Name |
|
| Description | ChIPAb+ CTCF - ChIP Validated Antibody and Primer Set |
| Overview | All 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 |
|
| Background Information | CTCF, 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. |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
|
| Presentation | Anti-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 Level | MQ100 |
| Applications | |
|---|---|
| Application | The 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 |
|
| Application Notes | Chromatin 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 | |
|---|---|
| Immunogen | His-tagged recombinant protein corresponding to human CTCF. |
| Clone | 1.2.1.5.4 |
| Host | Mouse |
| Specificity | Recognizes CTCF, Mr ~130 kDa. |
| Isotype | IgG1κ |
| Species Reactivity |
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| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | This 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 |
|
| Purification Method | Protein G Purified |
| UniProt Number |
|
| UniProt Summary | Function: 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. |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Chromatin 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 |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable 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 Size | 25 assays |
| Material Package | 25 assays per set. Recommended use: ~2 μg of antibody per chromatin immunoprecipitation (dependent upon biological context). |