CheckMate™/Flexi® Vector Mammalian Two-Hybrid System

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Study Mammalian Protein Interactions, Expression and Post-Translational Modifications

  • Confirm, validate and study suspected protein or domain interactions
  • Generate stable cell lines for cell-based assays
  • Enables mammalian protein expression and post-translational modification studies in situ


Catalog number selected: C9360

$ 937.00
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CheckMate™/Flexi® Vector Mammalian Two-Hybrid System
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$ 937.00
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The CheckMate™/Flexi® System is designed to confirm, validate and study suspected interactions between two proteins or domains. This system can also be used to generate stable cell lines for cell-based assays. The system can allow mammalian protein expression and post-translational modification studies in an environment mimicking the native cell milieu. The CheckMate™/Flexi® System 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.

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.

The plasmids are available  and can be ordered separately from the system.

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


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

GenBank® Accession Number and Sequence Information

pGL4.31[luc2P/Gal4UAS/Hygro], DQ487213.


What's in the box?

Item Part # Size Available Separately

pFN10A (ACT) Flexi® Vector

C933B 1 × 5μg View Product

pFN11A (BIND) Flexi® Vector

C934B 1 × 5μg View Product

pGL4.31[luc2P/Gal4UAS/Hygro] Vector

C935A 1 × 20μg View Product

pBIND Vector

E245A 1 × 20μg View Product

pACT Vector

E246A 1 × 20μg View Product

pBIND-Id Control Vector

E247A 1 × 20μg View Product

pACT-MyoD Control Vector

E248A 1 × 20μg View Product

Certificate of Analysis

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Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions


Patents and Disclaimers

BY USE OF THIS PRODUCT, RESEARCHER AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE. If the researcher is not willing to accept the terms of this label license, 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 money 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 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 label license shall be governed under the laws of the State of Wisconsin, USA. This label license relates to Promega patents and/or patent applications on improvements to the luciferase gene.

U.S. Pat. Nos. 8,293,503, 9,018,014 and 8,367,403, European Pat. No. 1685247 and other patents and patents pending.

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.

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