TCF Reporter Plasmid Kit

Description
Catalogue Number	17-285
Brand Family	Upstate
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.
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.

Description

Catalogue Number

17-285

Brand Family

Upstate

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.

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.

Product Information
Components	FOPflash (mutant TCF binding sites) (Cat.# 21-169) TOPflash (TCF Reporter Plasmid) (Cat.# 21-170)
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.

Product Information

Components

FOPflash (mutant TCF binding sites) (Cat.# 21-169)
TOPflash (TCF Reporter Plasmid) (Cat.# 21-170)

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.

Applications
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

Applications

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	NM_000545.4
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	TCF1 MODY3 HNF1 HNF-1A TCF-1 HNF1A LFB1 HNF1a
UniProt Number	P20823 P15884 P15923
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. 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.

Biological Information

Entrez Gene Number

NM_000545.4

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

TCF1
MODY3
HNF1
HNF-1A
TCF-1
HNF1A
LFB1
HNF1a

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.
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.

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.

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.

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

Packaging Information

Material Size

1 kit

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

实验服务

办公用品

TCF Reporter Plasmid Kit

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