Description
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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.
Supplements – Forming disulfide bonds –
These are supplements for PUREfrex® series reconstituted coupled transcription/translation systems to promote synthesis of functional proteins containing disulfide bonds.
