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PURExpress

PURExpress® (NEB #E6800) is a reconstituted protein synthesis system based on the PUREsystem™ (Shimizu et al., 2001) where all necessary components needed for in vitro transcription and translation are purified from E. coli.

  • Defined system with all His-tagged proteins for coupled transcription/translation; Ribosome is not His-tagged
  • T7 RNA Polymerase drives in vitro transcription
  • Minimal nuclease and protease activity for stability of synthesized protein and encoding target
  • Templates can be either plasmid DNA, linear DNA or mRNA
  • Protein of interest can be synthesized and visualized in a few hours
  • Synthesizes various target peptides and proteins
  • Synthesized protein can be co-translationally radiolabeled or fluorescently labeled
  • Protein can be reverse-purified or subject to direct functional analysis
  • Applications include high throughput screening/directed evolution, synthetic biology, toxic or difficult to express protein synthesis, studies on protein folding, activity and protein-protein interactions
  • Due to reconstituted nature, several kits are offered where translation factors or macromolecules have been omitted to facilitate specific studies; PURExpress® ∆ (aa, tRNA) Kit (NEB #E6840), PURExpress® ∆ RF123 Kit (NEB #E6850), PURExpress® ΔRibosome Kit (NEB #E3313)
  • Compatible with the PURExpress Disulfide Bond Enhancer (NEB #E6820)

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PURExpress
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E. coli Ribosome

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PURExpress® Disulfide Bond Enhancer

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PURExpress® In Vitro Protein Synthesis Kit

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PURExpress® Δ (aa, tRNA) Kit

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PURExpress® Δ RF123 Kit

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PURExpress® Δ Ribosome Kit

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RNase Inhibitor, Murine

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FAQs for PURExpress
Protocols for PURExpress
Application Notes for PURExpress
The PURExpress® System Kit Components
  • His-Tagged Translation Factors
    • Initiation Factors (IF1, IF2, IF3)
    • Elongation Factors (EF-Tu, EF-Ts, EF-G)
    • Release Factors (RF1, RF2, RF3)
    • Ribosome Recycling Factor
    • 20 Aminoacyl tRNA synthetases
    • Methionyl tRNA formyltransferase
  • E. coli Ribosomes
  • E. coli tRNAs
  • Energy Regeneration System
  • NTPs, Amino Acids, Salts, Buffer

In addition, recombinant T7 RNA polymerase is used to couple transcription to translation. The PURE system represents an important step towards a totally defined in vitro transcription/translation system, thus avoiding the “black box” nature of the cell extract-based systems.

Protein Expression Using the PURExpress® In Vitro Protein Synthesis Kit
25 µl reactions containing 250 ng template DNA were incubated at 37°C for 2 hours. 2.5 µl of each reaction was analyzed by SDS-PAGE using a 10–20% Tris-glycine gel. Note that proteins can be purified using reverse affinity chromatography (reagents not supplied). The red dot indicates the protein of interest. Marker M is the Protein Ladder (NEB #P7703)
PUREXPRESS® is a registered trademark of New England Biolabs, Inc..
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Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. All other trademarks are the property of their respective owners. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

 


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