17-285 | TCF Reporter Plasmid Kit

FOPflash and TOPflash TCF reporter plasmids
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      Replacement Information
      Catalogue Number 17-285
      Brand Family Upstate
      Synonyms FOPFlash plasmid TOPFlash/FOPFlash TOPFlash plasmid
      Trade Name
      • Upstate
      Description TCF Reporter Plasmid Kit
      Overview 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.

      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.
      Product Information
      • TOPflash (TCF Reporter Plasmid) (Cat.# 21-170)
      • FOPflash (mutant TCF binding sites) (Cat.# 21-169)
      Presentation 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.
      Application Set 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 Summary 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.
      Gene Symbol
      • HNF1a
      • TCF1
      • MODY3
      • HNF1
      • HNF-1A
      • TCF-1
      • HNF1A
      • LFB1
      UniProt Number
      UniProt Summary 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.
      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 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.
      Packaging Information
      Material Size 1 kit
      Transport Information
      Supplemental Information


      Certificates of Analysis

      TitleLot Number
      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
      TCF Reporter Plasmid Kit - 2055312 2055312
      TCF Reporter Plasmid Kit - 21254 21254
      TCF Reporter Plasmid Kit - 2189962 2189962
      TCF Reporter Plasmid Kit - 22722 22722
      TCF Reporter Plasmid Kit - 2329807 2329807
      TCF Reporter Plasmid Kit - 23447 23447
      TCF Reporter Plasmid Kit - 2512496 2512496
      TCF Reporter Plasmid Kit - 27340 27340
      TCF Reporter Plasmid Kit - 28039 28039
      TCF Reporter Plasmid Kit - 32612 32612
      TCF Reporter Plasmid Kit - DAM1479561 DAM1479561
      TCF Reporter Plasmid Kit - DAM1541077 DAM1541077
      TCF Reporter Plasmid Kit - DAM1574299 DAM1574299
      TCF Reporter Plasmid Kit - DAM1658630 DAM1658630
      TCF Reporter Plasmid Kit - DAM1718525 DAM1718525
      TCF Reporter Plasmid Kit - DAM1791460 DAM1791460
      TCF Reporter Plasmid Kit - JBC1362522 JBC1362522
      TCF Reporter Plasmid Kit - JBC1903606 JBC1903606
      TCF Reporter Plasmid Kit -2489083 2489083


      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

      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

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

      Show Abstract
      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


      Related Products & Applications

      Kit Component

       Catalogue 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