PURExpress® In Vitro Protein Synthesis Kit

PURExpress® 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
  • Minimal nuclease and protease activity
  • Protein of interest can be synthesized and visualized in a few hours
  • Simple protocol and easy setup with Solutions A and B
  • Templates can be either plasmid DNA, linear DNA or mRNA
  • 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 (see companion products)

Ordering Information

E6800S_pdp_800X450
  • 10 reactions
    $245.00
  • 100 reactions
    $2,160.00
  • Product Information
    A rapid method for gene expression analysis, PURExpress® is a novel cell-free transcription/translation system reconstituted from the purified components necessary for E. coli translation. The relative nuclease-free and protease-free nature of the PURExpress platform preserves the integrity of DNA and RNA templates/ complexes and results in proteins that are free of modification and degradation. Transcription and translation are carried out in a one-step reaction, and require the mixing of only two tubes. With results available in a few hours, PURExpress saves valuable laboratory time and is ideal for high throughput technologies.

    PURExpress Citations

    Figure 1: Protein expression using the PURExpress® In Vitro Protein Synthesis Kit 
    25 μl reactions containing 250 ng template DNA and 20 units RNase Inhibitor 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. The red dot indicates the protein of interest. Marker M is the Protein Ladder (NEB #P7703 ).
    Figure 2: Incorporation of 35S-methionine enables visualizationof protein by autoradiography Figure 2: Incorporation of 35S-methionine enables visualizationof protein by autoradiography
    25 μl reactions containing 250 ng template DNA, 20 units RNase Inhibitor and 2 μl 35S-met were incubated at 37°C for 2 hours. 2.5 μl of each reaction was analyzed by SDS-PAGE, the gel was fixed for 10 minutes, dried for 2 hours at 80°C and exposed to x-ray film for 5 hours at -80°C.
    Figure 3: Schematic diagram of protein synthesis and purification by PURExpressFigure 3: Schematic diagram of protein synthesis and purification by PURExpress

    Figure 4: Expression and reverse purification of DHFR (A) and T4 DNA Ligase (B) using PURExpress Figure 4: Expression and reverse purification of DHFR (A) and T4 DNA Ligase (B) using PURExpress
    125 μl reactions were carried out according to recommendations in the accompanying manual. Samples were analyzed on a 10–20% Tris-glycine gel and stained with Coomassie Blue. Note that in both cases, the desired protein can be visualized in the total protein fraction. The red dot indicates the protein of interest. Marker M is the Protein Ladder (NEB #P7703 ).

    Kit Components

    The following reagents are supplied with this product:

    Store at (°C) Concentration
    PURExpress Solution A -80
    Control (DHFR) template (10 μl) -80
    Solution B (75 μl) -80 3.3 X
    Product Categories:
    Cell-Free Expression Products
    Applications:
    Cell-Free Protein Expression,
    Toxic Protein Expression,
    Disulfide-Bonded Protein Expression
    • Advantages and Features
    • Properties and Usage
    • Related Products
    • Notes
    • References
  • FAQs & Tech Tips
  • Protocols & Manuals
  • Other Tools & Resources
  • Quality & Safety
  • Legal Information
    • Legal And Disclaimers
Loading Spinner