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PUREfrex™: Cell-free protein production system

PUREfrex™: Cell-free protein production system

PUREfrex™* is the purest cell-free protein synthesis kit based on the PURE (Protein Synthesis Using Recombinant Elements) system developed by Professor Takuya Ueda (University of Tokyo). The PURE system successfully reconstituted the E. coli transcription/translation process in the test tube.

PUREfrex™ applications

  • Easy assessment of the influence of mutations on a gene product.
  • Easy production of proteins which are toxic or prone to proteolytic degradation.
  • Fast identification of functional proteins from a pool of variants in multiwell plate format.
  • Quickest way to obtain proteins for:
    • Structural and kinetic studies
    • Antibody generation
    • Screening assays

PUREfrex™ benefits

PURE: Reducted level of contamination

PUREfrex™ is advanced in the purity by improving the preparation method of ribosomes, tRNAs and all protein constituents compared to the original PURE system. As a result, the contaminating lipopolysaccharide [LPS] from E.coli is reduced below 0.1EU per µL of reaction [Fig.1], and other contaminants like RNase and ß-galactosidase are also reduced. Therefore, PUREfrex™ is perfectly adapted to any downstream application.

PUREfrex™ contamination level

Fig.1 Lipopolysaccharides [LPS] activity detected in reaction samples using the PUREfrex™ kit vs competitor kits. The contaminating LPS activity detected in the PUREfrex™ sample is below 0.1 EU/µL.



EASY: Any tag for your protein purification

Since PUREfrex™ protein components are free of tag, you can fuse your protein of interest to any tag to ease purification step [Fig.2].

His-tagged GFP production using PUREfrex

Fig.2: His-tagged GFP production using PUREfrex™.
1: Synthesis reaction sample
2: Flowthrough sample
3: Washing step sample
4: Purified His-tagged GFP sample

FAST: Quick protein production and purification

With PUREfrex™, obtain the expression product of a genetic sequence only takes 2-4 hours.

FLEXIBLE: Synthesis of various proteins

Because PUREfrex™ only contains fully characterized components, it allows an easy adjustment of the assay conditions. With PUREfrex™, Eukaryotic and Prokaryotic proteins can be easily produced [Fig.3] from plasmid or PCR templates.

SDS-PAGE of Eukaryotic and Prokaryotic protein samples produced using PUREfrex

Fig.3 SDS-PAGE of Eukaryotic* and Prokaryotic** protein samples produced using PUREfrex™ : -DNA= negative control, GFP*= Green fluorescent protein, DHFR** = Dihydrofolate reductase, GST* = Glutathione S-transferase, ß-Gal** = ß-Galactosidase, bR** = Bacteriorhodopsin, Luc* = Luciferase, CS* = Citrate synthase, MDH* = Malate dehydrogenase, Her2d1* = Domain1 of Her2

PUREfrex™ technology



Purefrex technology

PUREfrex™ system includes highly purifi ed factors [ref.1, 2] necessary for:

  1. Transcription: RNA Polymerase + NTPs
  2. Aminoacylation: Aminoacyl-tRNA synthetase + tRNA + aa
  3. Translation: Ribosome + Initiation factors + Elongation factors + Release factors + RRF
  4. Energy Regeneration: CK, NDK, ADK, PPiase
 
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*PUREfrex™ is developed for in vitro research use only. PUREfrex™ should not be used for the therapy, diagnostic or administration to animals including human and should not be used as food or cosmetics etc.

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