2013 Promega catalogue

Life
Science
Catalog
2013
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Cell Signaling
Prokaryotic Cell-Free Protein
Expression
S30 T7 High-Yield Protein Expression
System
Product Size Cat.# Price ($)
S30 T7 High-Yield Protein Expression System 24 reactions L1110 659
8 reactions L1115 261
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: The E. coli S30 T7 High-Yield Protein Expression System is
designed to express up to 500μg/ml of protein in 1 hour from plasmid vectors
containing a T7 promoter and a ribosome binding site. The protein expression
system provides an extract that contains T7 RNA polymerase for transcription
and is deficient in OmpT endoproteinase and lon protease activity. All other
necessary components in the system are optimized for protein expression.
This results in greater stability and enhanced expression of target proteins.
Features:
• Obtain Data Faster: Protein expression in only one hour, not days as with
cell-based expression.
• Complete System: No requirement to purchase additional reagents.
• Achieve High Protein Expression: Express up to 500μg/ml of protein
for multiple applications.
• Scalable: Convenient screening protocol for high-throughput protein
expression.
• Flexible: Detect expressed proteins by Coomassie ® staining or
incorporation of a fluorescence or biotinylated modified tRNA.
Storage Conditions: Store at –70°C.
Protocol
S30 T7 High-Yield Protein Expression System Technical Manual
E. coli T7 S30 Extract System for Circular
DNA
Part#
TM306
Product Size Cat.# Price ($)
E. coli T7 S30 Extract System for Circular DNA 30 reactions L1130 580
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
Part#
E. coli T7 S30 Extract System for Circular DNA Technical Bulletin TB219
E. coli S30 Extract System for Linear
Templates
Product Size Cat.# Price ($)
E. coli S30 Extract System for Linear Templates 30 reactions L1030 544
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-P R ). 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 templates can be used: 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: System 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
Part#
E. coli S30 Extract System for Linear Templates Technical Bulletin TB102
Section
Contents
Table of
Contents
264
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