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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| H, M | WB, DB, ChIP |
| Description | |
|---|---|
| Catalogue Number | 17-10109 |
| Brand Family | Upstate |
| Trade Name |
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| Description | ChIPAb+ TCF-4 - 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+ TCF-4 set includes the TCF-4 antibody, a negative control normal mouse IgG, and qPCR primers which amplify a 198 bp region of human SP5 promoter. The TCF-4 and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of TCF-4-associated chromatin. |
| Alternate Names |
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| Background Information | TCF4, also known as TCF7L2, is expressed widely during development. Gene targeting study indicates that it is required to maintain the crypt stem cells of the small intestine. TCF4 has many different splicing isoforms and they are expressed differentially in tissues and in cancers of different stages. Studies also indicate that variant of the TCF4 gene confers an increased risk of type 2 diabetes. |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
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| Presentation | Anti-TCF-4 (mouse monoclonal). One vial containing 50 µg of purified mouse monoclonal IgG2a in 50 µL of buffer containing 0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl, 0.05% sodium azide and 30% glycerol. Store at -20°C. Normal Mouse IgG. Two vials containing 25 µg of purified IgG in 25 µL of storage buffer containing 0.1% sodium azide. Store at -20°C. ChIP Primers, SP5 promote. One vial containing 75 μL of 5 μM of each primer specific for human SP5 promotor. Store at -20°C. FOR: GGG TCT CCA GGC GGC AAG REV: AGC GAA AGC AAA TCC TTT GAA TCC |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | This ChIPAb+ TCF-4 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. |
| Key Applications |
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| Application Notes | Chromatin Immunoprecipitation: Representative lot data. Sonicated chromatin prepared from HT29 cells (3 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of Normal Mouse IgG or 2 µg Anti-TCF-4 and the Magna ChIP™ G Kit (Cat. # 17-611). Successful immunoprecipitation of TCF-4 associated DNA fragments was verified by qPCR using ChIP Primers, SP5 Promotor as a positive locus, and GAPDH promoter primers as a negative locus (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™ G (Cat. # 17-409) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details. Western Blot Analysis: Representative lot data. Cos-1 cells were transfected with TCF-4 cDNA in pUSE (lane 2) or pUSE (lane 1). RIPA lysates were prepared, resolved by electrophoresis, transferred to nitrocellulose and probed with anti-TCF-4 (1 mg/mL). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates TCF-4 (~66 kDa) (Please see figures). Immunocytochemistry/Immunohistochemistry: A previous lot of this antibody is reported to show positive immunostaining for TCF-4 in cells fixed with 4% paraformaldehyde/PBS (Amir, A. L., 2003). Immunoprecipitation: A previous lot of this antibody was reported to immunoprecipitate TCF-4 from SW620 whole cell lysate as a complex with Beta-Catenin (Persad, S., 2001). Supershift Assay (EMSA): A previous lot of this antibody was reported to supershift TCF-4/DNA complexes in electrophoretic mobility shift assays (EMSA) (Amir, A. L., 2003). |
| Biological Information | |
|---|---|
| Immunogen | Amino acids 31-331 of human TCF-4. |
| Host | Mouse |
| Specificity | This antibody recognizes TCF-4 |
| Isotype | IgG2a |
| Species Reactivity |
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| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | SOX-TCF_HMG-box, class I member of the HMG-box superfamily of DNA-binding proteins. These proteins contain a single HMG box, and bind the minor groove of DNA in a highly sequence-specific manner. Members include SRY and its homologs in insects and vertebrates, and transcription factor-like proteins, TCF-1, -3, -4, and LEF-1. They appear to bind the minor groove of the A/T C A A A G/C-motif. |
| Gene Symbol |
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| Purification Method | Protein G Chromatography |
| UniProt Number |
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| UniProt Summary | FUNCTION: Participates in the Wnt signaling pathway and modulates MYC expression by binding to its promoter in a sequence-specific manner. Acts as repressor in the absence of CTNNB1, and as activator in its presence. Activates transcription from promoters with several copies of the Tcf motif CCTTTGATC in the presence of CTNNB1. TLE1, TLE2, TLE3 and TLE4 repress transactivation mediated by TCF7L2 and CTNNB1. Expression of dominant-negative mutants results in cell-cycle arrest in G1 (By similarity). Necessary for the maintenance of the epithelial stem-cell compartment of the small intestine. SIZE: 619 amino acids; 67919 Da SUBUNIT: Interacts with TGFB1I1. Interacts with CTNNB1 (via the armadillo repeat); forms stable transcription complex. Interacts with EP300 (By similarity). SUBCELLULAR LOCATION: Nucleus. Note=Diffuse pattern. Colocalizes with SUMO1 and PIAS4 in a subset of PML (promyelocytic leukemia) nuclear bodies (By similarity). TISSUE SPECIFICITY: Detected in epithelium from small intestine, with the highest expression at the top of the crypts and a gradient of expression from crypt to villus. Detected in colon epithelium and colon cancer, and in epithelium from mammary gland and carcinomas derived therefrom. DISEASE: Constitutive activation and subsequent transactivation of target genes may lead to the maintenance of stem-cell characteristics (cycling and longevity) in cells that should normally undergo terminal differentiation and constitute the primary transforming event in colorectal cancer (CRC). SIMILARITY: Contains 1 HMG box DNA-binding domain. |
| Molecular Weight | ~66 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Chromatin Immunoprecipitation: Sonicated chromatin prepared from HT29 cells (3 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of Normal Mouse IgG or 2 µg of Anti-TCF-4 and the Magna ChIP™ G Kit (Cat. # 17-611). Successful immunoprecipitation of TCF-4 associated DNA fragments was verified by qPCR using ChIP Primers, SP5 Promotor (Please see figures). Please refer to the EZ-Magna ChIP™ G (Cat. # 17-409) 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: Variabillity 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). |