PUREfrex Product Pages

PUREfrex Product Manual and Package Inserts

Key Technical Notes

PUREfrex FAQ

PUREfrex in the Literature

Manufacturer's Site Links

Safety Data Sheets

Posters

  • General Protein Synthesis
    • Improvement of translational efficiency by N-terminal codon optimization in the reconstituted cell-free protein synthesis system [2016]
      •  
    • Improvement of the translation efficiency of proline residue-containing proteins using the reconstituted cell-free protein synthesis system (PUREfrex) [2018] VIEW
      •  
    • Investigation on how to synthesize active proteins by using a reconstituted cell-free protein synthesis system (PUREfrex) [2020]
      •  
  • Disulfide Bonding
    •  
    • Synthesis of proteins containing disulfide bonds using a reconstituted cell-free protein synthesis system (PUREfrex). [2019]
      •  
    • Synthesis of proteins containing disulfide bonds using PUREfrex® with human protein disulfide isomerase [2018] VIEW
      •  
    • Synthesis of functionally active proteins containing disulfide bonds using the new PURE system (PUREfrex) [2011]
    •  
  • Synthesis of Immunoglobulins and Biologics
    •  
    • Unique Cell-Free Protein Synthesis System, PUREfrex -Useful Platform for Protein Expression in the Development of Biologics- [2018] VIEW
      •  
    • Preparation of labeled cyclic peptides with PUREfrex [2017]
      •  
    • Development of a method for the synthesis of aglycosylated full-length IgG using PUREfrex [2017]
      •  
    • Efficient in vitro expression of aglycosylated full-length IgG using a reconstituted cell-free protein synthesis system, PUREfrex 2.0 [2016]
      •  
    • In vitro production of Fab, Toxin, and Immunotoxin using PUREfrex®2.0 with increased productivity [2015]
      •  
    • Synthesis of functionally active proteins containing disulfide bonds using the new PURE system (PUREfrex) [2011]

state-of-the-art protein synthesis by

PUREfrex® supplements expand the potential of base kits.

yyyy Molecular Chaperone Supplements Promote Protein Solubility and Protein Folding
Molecular chaperones such as hsp70 (e.g. E. coli DnaK) assist the correct folding of the newly synthesized proteins. Because base PUREfrex® kits do not contain molecular chaperones, some proteins may have difficulty forming the correct conformation and be insoluble after synthesis. In such cases, molecular chaperones could contribute to make them soluble and active.

Chaperone Supplement

DnaK Mix comprises highly purified and ratio-optimized E. coli DnaK, DnaJ and GrpE. DnaK, known as Hsp70, has ATPase activity and is stimulated by co-chaperones DnaJ and GrpE. DnaJ facilitates DnaK ATPase activity and binds to hydrophobic regions of synthesized proteins. GrpE stimulates ADP/ATP exchange of DnaK. DnaK Mix works well with PUREfrex®series and DsbC Set.

Reagent Qty Description Storage
DnaK Mix 25 ul  100µM DnaK, 20µM DnaJ and 20µM GrpE from E.coli in 30% glycerol buffer (No tags)  -80 C
 Dilution Buffer 500 ul  30% glycerol buffer   -20 C

Chaperone Supplement

GroE Mix comprises highly purified and ratio-optimized E. coli GroEL (known as Hsp60) and GroES (works in conjunction with GroEL). 

  

Reagent Qty Description Storage
 GroE Mix 12.5 ul   20µM GroEL, 14-mer form, and 40µM GroES, 7-mer form, from E.coli in 30% glycerol buffer (No tags)  -80 C
 Dilution Buffer 500 ul  30% glycerol buffer   -20 C

 

Redox Environment Supplements Promote Disulfide Bond Formation
Formation of disulfide bonds is important for folding and stability of secretory proteins such as enzymes or antibodies. Disulfide bonds are usually formed by oxidation of sulfhydryl groups (SH-) of adjacent cysteine residues. Therefore, disulfide bond formation efficiency depends on redox state. Additionally, disulfide bond isomerase which can catalyze the exchange of disulfide bridges may be required for correct cysteine pairing.

Redox Supplement

DsbC Set comprises highly purified E. coli DsbC (a disulfide bond isomerase) which can catalyze disulfide bridge exchange, and GSSG (oxidized Glutathione) to enforce an oxidized environment.

 

Reagent Qty Description Storage
GSSG 25 ul  Oxidized glutathione (60 mM) -20 C
DSBC 25 ul  E.coli DsbC (320 µM *2) (No tags)  -80 C
Dilution Buffer  500ul   30% glycerol buffer -20 C 

Redox Supplement

PDI Set comprises oxidized glutathione (GSSG), human PDI (protein disulfide isomerase) and human Ero1α (ER oxidoreductin-1 to reoxidize PDI).

Reagent Qty Description Storage
GSSG 25 ul  Oxidized glutathione (60 mM) -20 C
 PDI 25 ul Human protein disulfide isomerase with no tags (200 µM) -80 C
 Ero1a 25 ul  Human ER oxidoreductin-1 to reoxidize PDI with no tags (5 µM) -80 C 
Dilution Buffer 500 ul 30% glycerol buffer -20 C

 

Elongation Factor P (EF-P) promotes translation of consecutive proline residues.
The E. coli translation factor EF-P (elongation factor P), a bacterial homolog of eukaryotic initiation factor 5A (eIF5A), functions to promote peptide bond formation between the first methionine and the next amino acid as well as the elongation of consecutive proline residues (3 or more residues). In bacteria, LYS34 of EF-P is modified post translationally to ß-lysllysine by EpmA (YjeA/GenX) and EpmB (YjeK). This modification is important to EF-P activity. PUREfrex EF-P Supplement contains this modification at LYS34.

Transcription Factor Supplement

 

EF-P Supplement comprises recombinant EF-P (modified at LYS34) and dilution buffer. Standard PUREfrex® kits do not contain EF-P. Addition of EF-P to PUREfrex reactions can improve the yield of proteins containing consecutive proline residues such as Pro-Pro-Pro and Pro-Pro-Gly by preventing ribosome stalling and promoting the formation of peptide bonds.

 

Reagent Qty Description Storage
EF-P 12.5 ul   Elongation Factor P (40µM) (No tags)  -80 C
Dilution Buffer 500 ul  30% glycerol buffer   -20 C

 

PUREfrex Ordering Information
Product Name
(click for price and order info)		 Catalog Number Total Reaction Volume Product Information Page
Kits
PUREfrex 2.0
 GFK-PF201-0.1-EX 100 ul 2.0
GFK-PF201-0.25-EX 250 ul
GFK-PF201-2ML-EX 1 x 2 ml
GFK-PF201-10ML-EX 5 x 2 ml
GFK-PF201-50ML-EX 2 x 25ml
Current generation kit for protein synthesis. Contains DTT as a reducing agent.
PUREfrex 2.1
 GFK-PF213-0.1-EX 100 ul 2.1
GFK-PF213-0.25-EX 250 ul
GFK-PF213-2ML-EX 1 x 2 ml
 GFK-PF213-10ML-EX  5 x 2 ml
 GFK-PF213-50ML-EX 2 x 25ml
Current generation kit for proteins with disulfide bonds. Formulated for user control of redox environment. 
PUREfrex 1.0
 GFK-PF001-0.25-EX 250 ul 1.0
GFK-PF001-2ML-EX 1 x 2 ml
 GFK-PF001-10ML-EX 5 x 2 ml
 GFK-PF001-50ML-EX 2 x 25ml
Prior generation kit for protein science research and synthetic biology. Components are fully disclosed.
Custom formulated kits are based on 1.0 components. Please inquire about custom kits.   
PUREfrex Reaction Supplements
DnaK Mix GFK-PF003-0.5-EX for 500 ul of reaction mix Supplements 
GFK-PF003-10ML-EX 5 x 2 ml
GFK-PF003-50ML-EX 2 x 25ml
GroE Mix GFK-PF004-0.5-EX for 500 ul of reaction mix Supplements 
PF004-10ML-EX 5 x 2 ml
PF004-50ML-EX 2 x 25ml
Supplements for synthesis of aggregate-prone proteins.
DsbC Set GFK-PF005-0.5-EX for 500 ul of reaction mix Supplements 
PF005-10ML-EX 5 x 2 ml
PF005-50ML-EX 2 x 25ml
PDI Set GFK-PF006-0.5-EX for 500 ul of reaction mix  Supplements 
PF006-10ML-EX 5 x 2 ml
PF006-50ML-EX 2 x 25ml
Supplements for synthesis of proteins containing disufide bonds.
EF-P GFK-PFS052-0.5-EX for 500 ul of reaction mix  Supplements 
PFS052-10ML-EX 5 x 2 ml
PFS052-50ML-EX 2 x 25ml
Supplement for use when consecutive prolines are in target protein.