PURExpress® Δ RF123 Kit

PURExpress® Δ RF123 kit is a variation of the PURExpress In vitro Protein Synthesis Kit where the release factors 1, 2 and 3 are omitted from the translation mix.

  • RF1, RF2 and RF3 are supplied separately to allow users to control protein synthesis with/without release factors of their choice
  • Essentially RF1 and RF3 free, RF2 is greatly reduced to minimal levels
  • Allows for robust recovery of activity-encoding nucleic acid template in library screening applications
  • Applications include non-natural amino acid incorporation, ribosome display and mRNA display

Ordering Information

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  • 10 reactions
    $350.00
  • 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. With minimal nuclease and protease activity, the PURExpress system 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. 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 PURExpress. Figure 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
    RF1 (10 μl) -80
    RF2 (10 μl) -80
    RF3 (10 μl) -80
    Product Categories:
    Cell-Free Expression Products
    Applications:
    Cell-Free Protein Expression
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