Cell Signaling Streptavidin Product Streptavidin For Laboratory Use. For additional information see page 10. Streptavidin Alkaline Phosphatase Product Streptavidin Alkaline Phosphatase For Laboratory Use. For additional information see page 10. Protein Interactions CheckMate™/Flexi® Vector Mammalian TwoHybrid System Product CheckMate™/Flexi® Vector Mammalian Two-Hybrid System Available Separately pFN10A (ACT) Flexi® Vector pFN11A (BIND) Flexi® Vector pGL4.31[luc2P/GAL4UAS/Hygro] Vector CheckMate™ Positive Control Vectors CheckMate™ Negative Control Vectors For Research Use Only. Not for Use in Diagnostic Procedures. 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. 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. Size Cat.# Price (Fr) 1 each C9360 1094.00 20 µg C9331 476.00 20 µg C9341 476.00 20 µg C9351 618.00 1 set C9370 268.00 1 set C9380 268.00 CheckMate™ Mammalian Two-Hybrid System Product Size Cat.# Price (Fr) CheckMate™ Mammalian Two-Hybrid System 1 system E2440 1021.00 For Research Use Only. Not for Use in Diagnostic Procedures. Description: Two-hybrid systems are extremely powerful methods for detecting protein:protein interactions in vivo. The basis of two-hybrid systems is the modular domains found in some transcription factors: a DNA-binding domain, which binds to a specific DNA sequence, and a transcriptional activation domain, which interacts with the basal transcriptional machinery. A transcriptional activation domain in association with a DNA-binding domain will promote the assembly of RNA polymerase II complexes at the TATA box and increase transcription. In the CheckMate™ Mammalian Two-Hybrid System the DNA-binding domain and the transcriptional activation domain, produced by separate plasmids, are closely associated when one protein (“X”) fused to a DNA-binding domain interacts with a second protein (“Y”) fused to a transcriptional activation domain. In this system, interaction between proteins X and Y results in transcription of a reporter gene. Features: • Mammalian 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. • Convenient Quantitation: The Dual-Luciferase® Reporter Assay System is used for detection. • Internal Control: Renilla luciferase normalizes transfection efficiency. • Fast Transient Assay: Results obtained two days after transfection, as compared to 3–4 days with the yeast system. • Stable Transfectants: The pACT Vector contains the neomycin phosphotransferase gene, which allows for selection of stable transfectants. Storage Conditions: Store at –20°C. Protocol CheckMate™ Mammalian Two-Hybrid System Technical Manual Part# TM049 Size Cat.# Price (Fr) 0.5 ml V5591 213.00 Size Cat.# Price (Fr) 1 mg Z7041 117.00 Features: • 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. Storage Conditions: Store at –20°C. Protocol CheckMate™/Flexi® Vector Mammalian Two-Hybrid System Technical Manual Part# TM283 14 For complete and up-to-date product information visit: www.promega.com/catalog 255 Protein Purification and Interactions Pagina 258
Pagina 260Heeft u een catalogus, i-brochure of web magazines? Gebruik Online Touch: PDF online op uw website plaatsen.
Promega Switzerland - Life Sciences Catalog 2012 Lees publicatie 100Home