PUREfrex 2.1

Cosmo Bio LTD

Catalog No: GFK-PF213-0.25-EX



Explore all PUREfrex® products and accessories from our PUREfrex® homepage.



PUREfrex^® 2.1 is the latest iteration of the PUREfrex^® series reconstituted coupled transcription/translation system that now makes it possible for user control over reaction redox conditions to promote high-yield of functional disulfide bond-containing proteins. It also features reduced RNase, β-galactosidase and LPS contamination and boosted protein yield compared to PUREfrex^® 1.0.

Formation of disulfide bond is an important process for folding and stability of extracellular and membrane proteins. Disulfide bonds are usually formed by oxidation of sulfhydryl groups (SH-) of adjacent cysteine residues. Thus, the efficiency of disulfide bond formation depends on redox state. Additionally, disulfide bond isomerase which catalyzes the exchange of disulfide bridges, may be required for correct cysteine pairing.

The redox state of PUREfrex^® series reconstituted coupled transcription/translation reactions reflects the nature of the reducing agent and the ratio of reducing and oxidizing agents. The reducing agent in PUREfrex^® 2.0(contained in Solution I) is DTT. By contrast, Solution I of PUREfrex^® 2.1 contains neither a reducing agent nor cysteine, making it possible to tailor the redox environment to optimize synthesis of your protein of interest by utilizing provided independent solutions of cysteine, DTT and reduced glutathione (GSH). Other reducing agents such as 2-mercaptoethanol (not provided) may also be used.

PUREfrex^® 2.1 enjoys the improvements incorporated into PUREfrex^® 2.0, including upgraded purification processes and an optimized composition that together reduce RNase, β-galactosidase and LPS contamination and boost protein yield 2-10 times compared to PUREfrex^® 1.0. All proteinaceous components of PUREfrex^® 2.1 are free of fusion tags, allowing users the freedom to incorporate any chosen tag for protein purification/detection.

For proteins whose functional conformation requires the assistance of molecular chaperones or, for further assistance in formation of disulfide bonds, we offer supplemental reagents that work in conjunction withPUREfrex^® 2.1.

Supplements – Chaperones –

These are supplements for PUREfrex^® series reconstituted coupled transcription/translation systems to promote production and solubility of aggregation-prone proteins.


• DnaK Mix (#PF003-0.5-EX)

• GroE Mix (#PF004-0.5-EX)


Supplements – Forming disulfide bonds –

>These are supplements for PUREfrex^® series reconstituted coupled transcription/translation systems to promote synthesis of functional proteins containing disulfide bonds.


• DsbC Set (#PF005-0.5-EX)

• PDI Set (#PF006-0.5-EX)






Citations for PUREfrex 2.1 – 18 Found

Curli synthesis and biofilm formation in enteric bacteria are controlled by a dynamic small RNA module made up of a pseudoknot assisted by an RNA chaperone.
Nucleic Acids Res. 2014 Apr;42(7):4682-96. doi: 10.1093/nar/gku098. Epub 2014 Feb 1. 
- Comments: Demonstrates the benefits of PUREfrex over PURExpress

Cell-free synthesis of the torque-generating membrane proteins, PomA and PomB, of the Na+-driven flagellar motor in Vibrio alginolyticus.
J Biochem. 2008 Nov;144(5):635-42. doi: 10.1093/jb/mvn110. Epub 2008 Sep 6. 
- Comments: Demonstrates the benefits of PUREfrex over PURExpress

RNA, but not protein partners, is directly responsible for translational silencing by a bacterial Hfq-binding small RNA.
Proc Natl Acad Sci U S A. 2008 Jul 29;105(30):10332-7. doi: 10.1073/pnas.0803106105. Epub 2008 Jul 23. 
- Comments: Demonstrates the benefits of PUREfrex over PURExpress

YaeJ is a novel ribosome-associated protein in Escherichia coli that can hydrolyze peptidyl-tRNA on stalled ribosomes.
Nucleic Acids Res. 2011 Mar;39(5):1739-48. doi: 10.1093/nar/gkq1097. Epub 2010 Nov 3. 
- Comments: Demonstrates the benefits of PUREfrex over PURExpress

Salmonella enterica serotype Typhimurium DT104 ArtA-dependent modification of pertussis toxin-sensitive G proteins in the presence of [32P]NAD.
Microbiology (Reading). 2009 Nov;155(Pt 11):3710-3718. doi: 10.1099/mic.0.028399-0. Epub 2009 Aug 20. 
- Comments: Demonstrates the benefits of PUREfrex over PURExpress

SmpB triggers GTP hydrolysis of elongation factor Tu on ribosomes by compensating for the lack of codon-anticodon interaction during trans-translation initiation.
J Biol Chem. 2006 Jun 9;281(23):15987-96. doi: 10.1074/jbc.M512165200. Epub 2006 Apr 6. 
- Comments: Demonstrates the benefits of PUREfrex over PURExpress

Roles of Tom70 in import of presequence-containing mitochondrial proteins.
J Biol Chem. 2009 Nov 13;284(46):31635-46. doi: 10.1074/jbc.M109.041756. Epub 2009 Sep 18. 
- Comments: Demonstrates the benefits of PUREfrex over PURExpress

In vitro genetic reconstruction of bacterial transcription initiation by coupled synthesis and detection of RNA polymerase holoenzyme.
Nucleic Acids Res. 2010 Jul;38(13):e141. doi: 10.1093/nar/gkq377. Epub 2010 May 10. 
- Comments: Demonstrates the benefits of PUREfrex over PURExpress

A small RNA regulates multiple ABC transporter mRNAs by targeting C/A-rich elements inside and upstream of ribosome-binding sites.
Genes Dev. 2007 Nov 1;21(21):2804-17. doi: 10.1101/gad.447207. 
- Comments: Demonstrates the benefits of PUREfrex over PURExpress

A multiprotein binding interface in an intrinsically disordered region of the tumor suppressor protein interferon regulatory factor-1.
J Biol Chem. 2011 Apr 22;286(16):14291-303. doi: 10.1074/jbc.M110.204602. Epub 2011 Jan 18. 
- Comments: Demonstrates the benefits of PUREfrex over PURExpress

Tom20 and Tom22 share the common signal recognition pathway in mitochondrial protein import.
J Biol Chem. 2008 Feb 15;283(7):3799-807. doi: 10.1074/jbc.M708339200. Epub 2007 Dec 6. 
- Comments: Demonstrates the benefits of PUREfrex over PURExpress

Cortisol is involved in temperature-dependent sex determination in the Japanese flounder.
Endocrinology. 2010 Aug;151(8):3900-8. doi: 10.1210/en.2010-0228. Epub 2010 Jun 9. 
- Comments: Demonstrates the benefits of PUREfrex over PURExpress

Role of antibiotic ligand in nascent peptide-dependent ribosome stalling.
Proc Natl Acad Sci U S A. 2011 Jun 28;108(26):10496-501. doi: 10.1073/pnas.1103474108. Epub 2011 Jun 13. 
- Comments: Demonstrates the benefits of PUREfrex over PURExpress

In vitro methods for peptide display and their applications.
Brief Funct Genomics. 2011 May;10(3):125-34. doi: 10.1093/bfgp/elr010. 
- Comments: Demonstrates the benefits of PUREfrex over PURExpress

Functional role played by the glycosylphosphatidylinositol anchor glycan of CD48 in interleukin-18-induced interferon-gamma production.
J Biol Chem. 2005 May 6;280(18):18056-62. doi: 10.1074/jbc.M413297200. Epub 2005 Mar 10. 
- Comments: Demonstrates the benefits of PUREfrex over PURExpress

Involvement of Hu and heterogeneous nuclear ribonucleoprotein K in neuronal differentiation through p21 mRNA post-transcriptional regulation.
J Biol Chem. 2005 Apr 1;280(13):12690-9. doi: 10.1074/jbc.M411119200. Epub 2005 Jan 25. 
- Comments: Demonstrates the benefits of PUREfrex over PURExpress

Curli synthesis and biofilm formation in enteric bacteria are controlled by a dynamic small RNA module made up of a pseudoknot assisted by an RNA chaperone.
Nucleic Acids Res. 2014 Apr;42(7):4682-96. doi: 10.1093/nar/gku098. Epub 2014 Feb 1. 
- Comments: Demonstrates the benefits of PUREfrex over PURExpress

Quantitative imaging of gene-expressing liposomes reveals rare favorable phenotypes.
Phys Biol. 2019 Apr 12;16(4):045002. doi: 10.1088/1478-3975/ab0c62. 
- Comments: Demonstrates the benefits of PUREfrex over PURExpress