Science Catalog 2012 Life Cell Signaling Eukaryotic Cell-Free Protein Expression TNT® T7 Insect Cell Extract Protein Expression System Product TN Size Cat.# Price (Fr) T® T7 Insect Cell Extract Protein Expression System pF25A ICE T7 Flexi® Vector pF25K ICE T7 Flexi® Vector For Research Use Only. Not for Use in Diagnostic Procedures. Description: The TNT® T7 Insect Cell Extract Protein Expression System is a convenient, quick, single-tube, coupled transcription/translation system for the cell-free expression of proteins. Protein synthesis reactions are initiated by the addition of a DNA template, eliminating the need for the time-consuming process of in vitro RNA synthesis. The extract is made from the commonly used Spodoptera frugiperda Sf21 cell line. All components necessary for the transcription/translation are present in the TNT® T7 ICE Master Mix. To initiate protein synthesis, the only component that must be added is the DNA template. Reactions are incubated at 28–30°C and are complete within 4 hours. Proteins are expressed from genes cloned downstream of the T7 promoter. Companion vectors have been designed to achieve optimal yield with this system (pF25A and pF25K). They contain untranslated region (UTR) sequences at the 5´ and 3´ ends of the gene coding region to enhance translation efficiency. Using the TNT® T7 Insect Cell Extract Protein Expression System and these vectors, 75μg/ml of functional protein can be produced. Features: • Obtain Data Faster: Protein is expressed in only 4 hours, not days as with cell-based expression. • Complete System: No requirement to purchase additional reagents. • Achieve High Protein Yields: Express up to 75μg/ml of protein for multiple applications. Storage Conditions: Store at –70°C. Protocol TNT® T7 Insect Cell Extract System Technical Manual pF25A ICE T7 Flexi® Vector pF25K ICE T7 Flexi® Vector Part# TM305 9PIL106 9PIL108 10 reactions L1101 327.00 40 reactions L1102 1145.00 20 µg L1061 399.00 20 µg L1081 399.00 TNT® SP6 High-Yield Wheat Germ Protein Expression System Product TN Size Cat.# Price (Fr) T® SP6 High-Yield Wheat Germ Protein Expression System Available Separately TN 40 reactions L3260 1184.00 10 reactions L3261 339.00 T® SP6 High-Yield Master Mix Minus Amino Acids For Research Use Only. Not for Use in Diagnostic Procedures. X808X Pls. Enq. Description: The TNT® SP6 High-Yield Wheat Germ Protein Expression System, based on an optimized wheat germ extract, is a single-tube, coupled transcription/translation system designed to express proteins in only two hours. Protein synthesized, in the range of 10–100μg/ml, can be used in multiple proteomic-based applications, as well as in high-throughput analysis. All components necessary for transcription/translation are provided in the extract, with the exception of the plasmid DNA or PCR template. Optional protein-labeling reagents must also be supplied by the user. For custom wheat germ extract (depleted amino acids), order Cat.# X808X (see Products, Available Separately). Features: • Save Time: You can generate protein in only two hours, as compared to days when using cell-based (E. coli) systems. • Choose Your Format: Use plasmid or PCR-generated templates to generate protein. • Achieve High Yields: Generate 10- to 20-fold more protein (10–100μg/ml) when compared to other cell-free systems. • Generate Usable Protein: Generate soluble, full-length protein and avoid problems associated with E. coli systems. Storage Conditions: Store at –70°C. Protocol TNT® SP6 High-Yield Wheat Germ Protein Expression System Technical Manual Part# TM282 224 For complete and up-to-date product information visit: www.promega.com/catalog Pagina 227
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