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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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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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.