PUREfrex®1.0

Kits

Product Catalog No. Quantity SDS Price (JPY) Instruction
PUREfrex®1.0 PF001-0.25-EX 250 µL for reaction US
EU

¥ 15000

PDF
PUREfrex®1.0 PF001-0.25-5-EX 1250 µL for reaction US
EU

¥67500

PDF

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https://www.cosmobiousa.com/

Overview

PUREfrex®1.0 is the kit for professional user who would like to customize the PUREfrex® component.
PUREfrex® is a reconstituted cell-free protein synthesis system based on the PURE system, which is reconstituted from individually purified factors involved in protein synthesis in E.coli. It has an advantage that the composition of the reagent can be easily customized according to the purpose.

Disclosure of Component Information

The component information of PUREfrex®1.0 is disclosed.
You may designed customize kits based on PUREfrex®1.0.

Custom Kits

Custom kits are made to order upon request.
Before considering a custom kit, it is recommended that you verify that the protein of interest can be synthesized with PUREfrex®1.0.
Please visit the web site for more information.
https://www.genefrontier.com/en/solutions/purefrex/lineup/custom-kit

Lower Contamination

By improving the purification process of components of PUREfrex®, contamination of RNase and β-galactosidase are greatly reduced, in addition to that, lipopolysaccharide (LPS) is also reduced to around 0.1 EU per 1 µL of reaction mixture. All proteinous components of PUREfrex® have no tags for purification and detection. It allows to fuse your protein with any tag to purify the product.

contamination test

* PURE system

Related Product

When your protein needs an assistance of molecular chaperone or a formation of a disulfide bond to form a correct conformation, we have supplemental reagents by simply adding to PUREfrex®1.0.

Supplements – Chaperones –

These are supplement for PUREfrex® which is molecular chaperone to be aggregate-prone protein in a soluble form.

DnaK Mix (#PF003-0.5-EX)
GroE Mix (#PF004-0.5-EX)

Supplements – Forming disulfide bonds –

This is supplement for synthesizing proteins containing disulfide bonds in an active form.

DsbC Set (#PF005-0.5-EX)
PDI Set (#PF006-0.5-EX)

Contents

Reagent Quantity Description Storage*1
Solution I 125 μL Amino acids, NTPs, tRNAs and substrates for enzymes etc. -20 °C
Solution II 12.5 μL Proteins in 30 % glycerol buffer -20 °C or -80 °C*2
Solution III 12.5 μL Ribosome (20 μM) -80 °C *2
DHFR DNA*3
(text)
10 μL PCR product containing a gene encoding dihydrofolate reductase (DHFR) from E.coli as a positive control. (20ng/μL) -20 °C

*1) Indicated temperature shows storage temperature after opening the kit. Please store the kit at -80 °C before opening.
*2) The rest of solution should be frozen rapidly in liquid nitrogen or dry ice/ethanol, and be stored at -80 °C. Divide into aliquots, if necessary, and avoid refreeze and thaw as much as possible.
*3) For synthesizing DHFR, add 1 µL of DHFR DNA to 20 µL of reaction.

Protocol

The amounts given are for 20 µL reaction. For scaling up the reaction, adjust the volume of reagents accordingly.

  1. Thaw Solution I by incubation at room temperature or 37 °C for 1 minute for completely dissolving, and then cool on ice.
  2. Thaw Solution II and III on ice.
  3. Mix Solution I, II and III respectively by vortex and centrifuge briefly to collect each solution at the bottom.
  4. Assemble the reaction mixture in a tube as follows. Add the template DNA to 1 – 3 ng/µL per 1 kb.
    Water 8-X μL
    Solution I 10 μL
    Solution II 1 μL
    Solution III 1 μL
    Template DNA X μL
    Total 20 μL
  5. Incubate the tube at 37 °C for 2 – 4 hours on a heat block or a water bath.
  6. Analyze the synthesized products. Please add the same amount of H2O to the reaction for the sample of SDS-PAGE.

Note

PUREfrex® is developed for in vitro research 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. To avoid the contamination of nuclease, nuclease-free-treated water, reagents and materials should be used. We also recommend wearing gloves and mask.

“PUREfrex® is Registered in U.S. Patent and Trademark Office”