Science Catalog 2012 Life Cell Signaling Prokaryotic Cell-Free Protein Expression Product S30 T7 High-Yield Protein Expression System Size Cat.# Price (Fr) S30 T7 High-Yield Protein Expression System 24 reactions L1110 755.00 8 reactions L1115 280.00 For Research Use Only. Not for Use in Diagnostic Procedures. Description: The E. coli S30 T7 High-Yield Protein Expression System is designed to express up to 500μg/ml of protein in 1 hour from plasmid vectors containing a T7 promoter and a ribosome binding site. The protein expression system provides an extract that contains T7 RNA polymerase for transcription and is deficient in OmpT endoproteinase and lon protease activity. All other necessary components in the system are optimized for protein expression. This results in greater stability and enhanced expression of target proteins. Features: • Obtain Data Faster: Protein expression in only one hour, not days as with cell-based expression. • Complete System: No requirement to purchase additional reagents. • Achieve High Protein Expression: Express up to 500μg/ml of protein for multiple applications. • Scalable: Convenient screening protocol for high-throughput protein expression. • Flexible: Detect expressed proteins by Coomassie® staining or incorporation of a fluorescence or biotinylated modified tRNA. Storage Conditions: Store at –70°C. Protocol S30 T7 High-Yield Protein Expression System Technical Manual E. coli T7 S30 Extract System for Circular DNA Product Size Cat.# Price (Fr) E. coli T7 S30 Extract System for Circular DNA 30 reactions L1130 755.00 For Research Use Only. Not for Use in Diagnostic Procedures. Description: The E. coli T7 S30 Extract System for Circular DNA simplifies the transcription/translation of DNA sequences cloned in plasmid or λ vectors containing a T7 promoter by providing an extract that contains T7 RNA polymerase for transcription and all components needed for translation. The investigator only supplies cloned DNA containing a T7 promoter and a ribosome binding site. This product is prepared by modifications of the method described by Zubay from an E. coli strain B deficient in OmpT endoproteinase and lon protease activity. This results in greater stability of expressed proteins that would otherwise be degraded by proteases if expressed in vivo. Features: • Flexible: Can translate using any clone that has a T7 promoter and a ribosome binding site. Other S30 extracts require an E. coli promoter. • Greater Stability: Reduced chance of expressed proteins degrading. • Complete: Contains all components needed for coupled transcription/ translation. • Low Background: Synthesizes very low levels of endogenous proteins. • Optimized: Premix is optimized for each lot of S30 Extract and contains all other required components (except amino acids), such as ribonucleotides, tRNAs, PEP (phosphoenol pyruvate) and salts. • Choose Your Configuration: Learn more about our custom options for this product at: www.promega.com/myway/ Storage Conditions: Store extract at –70°C. Check individual components for storage temperatures. Protocol E. coli T7 S30 Extract System for Circular DNA Technical Bulletin 230 Part# TB219 Part# TM306 E. coli S30 Extract System for Linear Templates Product E. coli S30 Extract System for Linear Templates For Research Use Only. Not for Use in Diagnostic Procedures. Description: The E. coli S30 Extract System for Linear Templates is prepared using minor modifications of the protocol described by Lesley and colleagues and allows successful transcription/translation of linear DNA templates. The investigator need only provide linear DNA containing a prokaryotic E. coli-like promoter (such as lacUV5, tac, λPL (con) and λ-PR). A ribosome binding site is required to direct the synthesis of proteins in vitro. In vitro-generated RNA from DNA templates lacking an E. coli promoter may also be used in this system, but protein yields will be decreased to 1–10% of that produced from linear DNA templates. Features: • Flexible: Many template choices: DNA fragments, PCR-synthesized DNA, ligated overlapping oligonucleotides, in vitro-generated RNA and prokaryotic RNA. • Greater Stability: Reduced chance of expressed proteins degrading. • Complete: Contains all necessary components for coupled transcription/ translation. • Low Background: Synthesizes very low levels of endogenous proteins. • Optimized: Premix is optimized for each lot of S30 Extract and contains all other required components (except amino acids), such as ribonucleotides, tRNAs, PEP (phosphoenol pyruvate) and salts. • Choose Your Configuration: Learn more about our custom options for this product at: www.promega.com/myway/ Storage Conditions: Store at –70°C. Protocol E. coli S30 Extract System for Linear Templates Technical Bulletin Part# TB102 E. coli S30 Extract System for Circular DNA Product Size Cat.# Price (Fr) E. coli S30 Extract System for Circular DNA 30 reactions L1020 719.00 For Research Use Only. Not for Use in Diagnostic Procedures. Description: The E. coli S30 Extract for Circular DNA simplifies the transcription/ translation of DNA sequences cloned in plasmid or λ vectors, providing a powerful tool for identifying and characterizing polypeptides. The investigator needs only to supply the cloned DNA containing the appropriate prokaryotic promoter and ribosome binding sites. The S30 Extract for Circular DNA Templates is prepared by modifications of the method described by Zubay from an E. coli strain B deficient in OmpT endoproteinase and lon protease activity. This results in a greater stability of expressed proteins that would be degraded by proteases if expressed in vivo. The S30 in vitro system also allows higher expression levels of proteins that are normally expressed at low levels in vivo due to the action of host-encoded repressors. Features: • Greater Stability: Reduced chance of expressed proteins degrading. • Complete: Contains all necessary components for coupled transcription/ translation. • Low Background: Synthesizes very low levels of endogenous proteins. • Optimized: Premix is optimized for each lot of S30 Extract and contains all other required components (except amino acids), such as ribonucleotides, tRNAs, PEP (phosphoenol pyruvate) and salts. • Choose Your Configuration: Learn more about our custom options for this product at: www.promega.com/myway/ Storage Conditions: Store at –70°C. Protocol E. coli S30 Extract System for Circular DNA Technical Bulletin Part# TB092 For complete and up-to-date product information visit: www.promega.com/catalog Size Cat.# Price (Fr) 30 reactions L1030 719.00 Pagina 233
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