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CheckMate/Flexi Vector Mammalian Two-Hybrid System (1 each)

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C9360

Description

The CheckMate™/Flexi® Vector Mammalian Two-Hybrid System provides a means to confirm, validate and study suspected interactions between two proteins or domains and can also be used to generate stable cell lines for cell-based assays. Developed primarily for mammalian proteins of interest, the system can allow protein expression and post-translational modifications in an environment mimicking the native cell milieu. It is patterned on the yeast two-hybrid system with one protein of interest ("X") fused to a DNA-binding domain and the other protein ("Y") fused to a transcriptional activation domain (Figure 1).

The system relies upon three plasmids that are co-transfected into mammalian cells, each plasmid having unique features. The pFN10A (ACT) Flexi® Vector contains a herpes simplex virus VP16 transcriptional activation domain upstream of the cloning site, and the pFN11A (BIND) Flexi® Vector contains the yeast GAL4DNA-binding domain upstream of the cloning site. The pFN11A (BIND) Flexi® Vector also expresses the Renilla reniformis luciferase under the control of the SV40 promoter, allowing normalization for differences in transfection efficiency. The third vector, pGL4.31[luc2P/GAL4UAS/Hygro] Vector, contains five GAL4 binding sites upstream of a minimal TATA box, which is upstream of a firefly luciferase gene that acts as a reporter for interactions between proteins X and Y.

This system differs from the original CheckMate™ Mammalian Two-Hybrid System in that the vectors are compatible with the Flexi® Vector System, which allows directional cloning and rapid, efficient and high-fidelity transfer of protein coding regions between a variety of Flexi® Vectors.

Features - Benefits

  • Mammalian-Based System: Interactions can be studied in the cell line of choice. Proteins are more likely to be in their native conformation. Post-translational modifications, such as glycosylation, phosphorylation and acylation, are better maintained.
  • Versatile: Vectors are based on the Flexi® Cloning technology, enabling convenient transfer of protein-coding regions for additional functional proteomics applications.
  • Convenient: The Dual-Luciferase® Reporter Assay System is used for detection.

Applications

  • Confirm, validate and study suspected interactions between two proteins or domains.

Specifications

GenBank®/EMBL Accession Number

pGL4.31[luc2P/GAL4UAS/Hygro], DQ487213.

Storage Conditions

Store at –20°C.

For product intended use please see Patents & Disclaimers tab.

Use Restrictions

C9360, C9331, C9341, C9351, C9370, C9380, C8640, L1001 For Research Use Only. Not for Use in Diagnostic Procedures.

Patents - Disclaimers

BY USE OF THIS PRODUCT, RESEARCHER AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE STATEMENT. If the researcher is not willing to accept the conditions of this limited use statement, and the product is unused, Promega will accept return of the unused product and provide the researcher with a full refund.
Researchers may use this product for research use only, no commercial use is allowed. Commercial Use means any and all uses of this product and derivatives by a party for monetary or other consideration and may include but is not limited to use in: (1) product manufacture; and (2) to provide a service, information or data; and/or resale of the product or its derivatives, whether or not such product or derivatives are resold for use in research. Researchers shall have no right to modify or otherwise create variations of the nucleotide sequence of the luciferase gene except that Researchers may: (1) create fused gene sequences provided that the coding sequence of the resulting luciferase gene has no more than four deoxynucleotides missing at the affected terminus compared to the intact luciferase gene sequence, and (2) insert and remove nucleic acid sequences in splicing research predicated on the inactivation or reconstitution of the luminescence of the encoded luciferase. No other use or transfer of this product or derivatives is authorized without the prior express written consent of Promega. In addition, Researchers must either: (1) use luminescent assay reagents purchased from Promega Corporation for all determinations of luminescence activity of this product and its derivatives; or (2) contact Promega to obtain a license for use of the product and its derivatives. Researchers may transfer derivatives to others for research use provided that at the time of transfer a copy of this label license is given to the recipients and recipients agree to be bound by the terms of this label license. With respect to any uses outside this label license, including any diagnostic, therapeutic or prophylactic uses, please contact Promega for supply and licensing information. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING FOR MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE WITH REGARDS TO THE PRODUCT. The terms of this agreement shall be governed under the laws of the State of Wisconsin, USA. The above license relates to Promega patents and/or patent applications on improvements to the luciferase gene.

U.S. Pat. No. 5,670,356.

Patent Pending.

For research use only. Persons wishing to use this product or its derivatives in other fields of use, including without limitation, commercial sale, diagnostics or therapeutics, should contact Promega Corporation for licensing information.

Licensed from University of Georgia Research Foundation, Inc., under U.S. Pat. Nos. 5,292,658, 5,418,155, Canadian Pat. No. 2,105,984 and related patents.

The method of recombinant expression of Coleoptera luciferase is covered by U.S. Pat. Nos. 5,583,024, 5,674,713 and 5,700,673. A license (from Promega for research reagent products and from The Regents of the University of California for all other fields) is needed for any commercial sale of nucleic acid contained within or derived from this product.

Patents for the foundational PCR process, European Pat. Nos. 201,184 and 200,362, expired on March 28, 2006. In the U.S., the patents covering the foundational PCR process expired on March 29, 2005.

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