17-285 | TCF Reporter Plasmid Kit

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      Replacement Information
      Catalogue Number17-285
      Brand Family Upstate
      Trade Name
      • Upstate
      DescriptionTCF Reporter Plasmid Kit
      OverviewFOPflash: 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 InformationThe 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.
      Product Information
      • FOPflash (mutant TCF binding sites) (Cat.# 21-169)
      • TOPflash (TCF Reporter Plasmid) (Cat.# 21-170)
      PresentationFOPflash, 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.
      ApplicationSet of transfection grade T cell factor (TCF) reporter plasmids for use in TOPFlash and FOPFlash wnt/b-catenin activity assays.
      Key Applications
      • Transfection
      Biological Information
      Entrez Gene Number
      Entrez Gene SummaryThis 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.
      Gene Symbol
      • TCF1
      • MODY3
      • HNF1
      • HNF-1A
      • TCF-1
      • HNF1A
      • LFB1
      • HNF1a
      UniProt Number
      UniProt SummaryFUNCTION: 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.
      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
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceRoutinely 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 ConditionsStable 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.
      Packaging Information
      Material Size1 kit
      Transport Information
      Supplemental Information




      Safety Data Sheet (SDS) 

      Certificates of Analysis

      TitleLot Number
      TCF Reporter Plasmid Kit 2893733
      TCF Reporter Plasmid Kit - 2433529 2433529
      TCF Reporter Plasmid Kit - 0609040122 0609040122
      TCF Reporter Plasmid Kit - 0703055812 0703055812
      TCF Reporter Plasmid Kit - 19030 19030
      TCF Reporter Plasmid Kit - 19495 19495
      TCF Reporter Plasmid Kit - 1984858 1984858
      TCF Reporter Plasmid Kit - 1999679 1999679
      TCF Reporter Plasmid Kit - 2003355 2003355
      TCF Reporter Plasmid Kit - 2014524 2014524


      Reference overviewPub Med ID
      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)  2000

      11033082 11033082
      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)  1999

      10221643 10221643
      The TAK1-NLK-MAPK-related pathway antagonizes signalling between beta-catenin and transcription factor TCF.
      Ishitani, T, et al.
      Nature, 399: 798-802 (1999)  1999

      Show Abstract
      10391247 10391247
      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

      Show Abstract
      9488439 9488439
      The Xenopus Wnt effector XTcf-3 interacts with Groucho-related transcriptional repressors.
      Roose, J, et al.
      Nature, 395: 608-12 (1998)  1998

      Show Abstract
      9783587 9783587
      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

      9118222 9118222

      Related Products & Applications

      Kit Component

      Catalog Number Description  
      21-169 FOPflash (mutant TCF binding sites) Show Pricing & Availability
      21-170 TOPflash (TCF Reporter Plasmid) Show Pricing & Availability


      Life Science Research > Genomic Analysis > Transfection and Protein Expression > Mammalian Expression > Mammalian Expression Vectors & Kits