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96-Well Plate
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48-602MAG
Buffer Detection Kit for Magnetic Beads
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17-285
Sigma-AldrichTCF Reporter Plasmid Kit
Set of transfection grade T cell factor (TCF) reporter plasmids for use in TOPFlash and FOPFlash wnt/b-catenin activity assays.
More>>Set of transfection grade T cell factor (TCF) reporter plasmids for use in TOPFlash and FOPFlash wnt/b-catenin activity assays. Less<<
TCF Reporter Plasmid Kit: SDB (Sicherheitsdatenblätter), Analysenzertifikate und Qualitätszertifikate, Dossiers, Broschüren und andere verfügbare Dokumente.
FOPflash: Transfection grade T cell factor (TCF) reporter plasmid containing two full and one incomplete copy of the TCF binding site (mutated) followed by three copies in the reverse orientation, upstream of the Thymidine Kinase (TK) minimal promoter and Luciferase open reading frame. This plasmid serves as a negative control to TOPflash which contains wild type TCF binding sites.
TOPflash: Transfection grade T-cell factor (TCF) reporter plasmid containing two sets (with the second set in the reverse orientation) of three copies of the TCF binding site (wild type) upstream of the Thymidine Kinase (TK) minimal promoter and Luciferase open reading frame. FOPflash containing mutated TCF binding sites is also available as a negative control.
Alternate Names
TOPFlash/FOPFlash
TOPFlash plasmid
FOPFlash plasmid
Background Information
The Wnt family encodes secreted glycoproteins implicated in cell growth regulation, cell fate determination, organogenesis and oncogenesis. Inferred from genetic analysis of wingless (wg) signaling in Drosophila, in mammalian cells, Wnts signal through the Frizzled family of receptors, to Dishevelled. Dishevelled down regulates glycogen synthase kinase-3ß (GSK-3ß) activity. This results in the stabilization and accumulation of cytosolic ß-catenin, a GSK-3ß substrate, which when phosphorylated by GSK-3ß, is targeted for ubiquitin-mediated proteolysis. ß-catenin then translocates to the nucleus, where in complexes with members of the Tcf/LEF family of transcription regulators, activates transcription of Tcf-responsive genes. This Tcf-reporter plasmid, along with FOPFLASH (Cat. # 21-169) enables quantitation of Wnt/Wg signaling in cells transfected with these constructs.
FOPflash, Catalog # 21-169, Lot # 30860, 5.5 kb. 5 μg affinity purified DNA eluted and aseptically packaged in 50 μL of 10 mM Tris-HCl, pH 8.0, 1 mM EDTA. Frozen solution.
TOPflash, Catalog # 21-170, Lot # 27752, 5.5 kb. 5 μg affinity purified DNA eluted and aseptically packaged in 50 μL of 10 mM Tris-HCl, pH 8.0, 1 mM EDTA. Frozen solution.
This gene encodes transcription factor 4, a basic helix-turn-helix transcription factor. The encoded protein recognizes an Ephrussi-box ('E-box') binding site ('CANNTG') - a motif first identified in immunoglobulin enhancers. This gene is expressed predominantly in pre-B-cells, although it is found in other tissues as well. Multiple alternatively spliced transcript variants that encode different proteins have been described.
FUNCTION: SwissProt: P20823 # Required for the expression of several liver specific genes. Binds to the inverted palindrome 5'-GTTAATNATTAAC-3'. SIZE: 631 amino acids; 67356 Da SUBUNIT: Binds DNA as a dimer. SUBCELLULAR LOCATION: Nucleus. TISSUE SPECIFICITY: Liver. DISEASE: SwissProt: P20823 # Defects in HNF1A may predispose to hepatic adenomas [MIM:142330]. Hepatic adenomas are benign tumors at risk of malignant transformation. Bi-allelic inactivation of HNF1A, whether sporadic or associated with MODY3, may be an early step in the developmant of some hepatocellular carcinomas. & Defects in HNF1A are the cause of maturity onset diabetes of the young type 3 (MODY3) [MIM:600496]; also symbolized MODY-3. MODY [MIM:606391] is a form of diabetes characterized by an autosomal dominant mode of inheritance, age of onset of 25 years or younger and a primary defect in insulin secretion. The clinical phenotype of MODY3 is characterized by severe insulin secretory defects, and by major hyperglycemia associated with microvascular complications. & Defects in HNF1A are a cause of susceptibility to insulin-dependent diabetes mellitus (IDDM) [MIM:222100]. SIMILARITY: SwissProt: P20823 ## Belongs to the HNF1 homeobox family. & Contains 1 homeobox DNA-binding domain.
Physicochemical Information
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Toxicological Information
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Safety Information
Product Usage Statements
Quality Assurance
Routinely evaluated by DNA Sequence Analysis to confirm the identity of TCF binding sites.
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 Conditions
Stable for 2 years at -20°C from date of shipment. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap.
Selective small molecule inhibitors of glycogen synthase kinase-3 modulate glycogen metabolism and gene transcription Coghlan, M P, et al Chem Biol, 7:793-803 (2000)
1999
Tcf-1-mediated transcription in T lymphocytes: differential role for glycogen synthase kinase-3 in fibroblasts and T cells Staal, F. J., et al Int Immunol, 11:317-23 (1999)
1998
The TAK1-NLK-MAPK-related pathway antagonizes signalling between beta-catenin and transcription factor TCF. Ishitani, T, et al. Nature, 399: 798-802 (1999)
1998
The Wnt signalling pathway regulates many developmental processes through a complex of beta-catenin and the T-cell factor/lymphoid enhancer factor (TCF/LEF) family of high-mobility-group transcription factors. Wnt stabilizes cytosolic beta-catenin, which then binds to TCF and activates gene transcription. This signalling cascade is conserved in vertebrates, Drosophila and Caenorhabditis elegans. In C. elegans, the proteins MOM-4 and LIT-1 regulate Wnt signalling to polarize responding cells during embryogenesis. MOM-4 and LIT-1 are homologous to TAK1 (a kinase activated by transforming growth factor-beta) mitogen-activated protein-kinase-kinase kinase (MAP3K) and MAP kinase (MAPK)-related NEMO-like kinase (NLK), respectively, in mammalian cells. These results raise the possibility that TAK1 and NLK are also involved in Wnt signalling in mammalian cells. Here we show that TAK1 activation stimulates NLK activity and downregulates transcriptional activation mediated by beta-catenin and TCF. Injection of NLK suppresses the induction of axis duplication by microinjected beta-catenin in Xenopus embryos. NLK phosphorylates TCF/LEF factors and inhibits the interaction of the beta-catenin-TCF complex with DNA. Thus, the TAK1-NLK-MAPK-like pathway negatively regulates the Wnt signalling pathway.
Two members of the Tcf family implicated in Wnt/beta-catenin signaling during embryogenesis in the mouse. Korinek, V, et al. Mol. Cell. Biol., 18: 1248-56 (1998)
1998
Tcf transcription factors interact with beta-catenin and Armadillo to mediate Wnt/Wingless signaling. We now report the characterization of genes encoding two murine members of the Tcf family, mTcf-3 and mTcf-4. mTcf-3 mRNA is ubiquitously present in embryonic day 6.5 (E6.5) mouse embryos but gradually disappears over the next 3 to 4 days. mTcf-4 expression occurs first at E10.5 and is restricted to di- and mesencephalon and the intestinal epithelium during embryogenesis. The mTcf-3 and mTcf-4 proteins bind a canonical Tcf DNA motif and can complex with the transcriptional coactivator beta-catenin. Overexpression of Wnt-1 in a mammary epithelial cell line leads to the formation of a nuclear complex between beta-catenin and Tcf proteins and to Tcf reporter gene transcription. These data demonstrate a direct link between Wnt stimulation and beta-catenin/Tcf transcriptional activation and imply a role for mTcf-3 and -4 in early Wnt-driven developmental decisions in the mouse embryo.
Tcf/Lef transcription factors mediate signalling from Wingless/Wnt proteins by recruiting Armadillo/beta-catenin as a transcriptional co-activator. However, studies of Drosophila, Xenopus and Caenorhabditis elegans have indicated that Tcf factors may also be transcriptional repressors. Here we show that Tcf factors physically interact with members of the Groucho family of transcriptional repressors. In transient transfection assays, the Xenopus Groucho homologue XGrg-4 inhibited activation of transcription of synthetic Tcf reporter genes. In contrast, the naturally truncated Groucho-family member XGrg-5 enhanced transcriptional activation. Injection of XGrg-4 into Xenopus embryos repressed transcription of Siamois and Xnr-3, endogenous targets of beta-catenin-Tcf. Dorsal injection of XGrg-4 had a ventralizing effect on Xenopus embryos. Secondary-axis formation induced by a dominant-positive Armadillo-Tcf fusion protein was inhibited by XGrg-4 and enhanced by XGrg-5. These data indicate that expression of Tcf target genes is regulated by a balance between Armadillo and Groucho.
Armadillo coactivates transcription driven by the product of the Drosophila segment polarity gene dTCF van de Wetering, M, et al Cell, 88:789-99 (1997)
1997
