Cube E.coli lysates for cell-free protein expression

In close collaboration with the group of Volker Doetsch and Frank Bernhard, Institute of Biophysics at the University of Frankfurt, Germany, we provide an open system that can be adapted for a range of applications, with dedicated protocols for membrane proteins, continous exchange (dialysis) reactions, and many more.  This system takes more work initially than ready to use kits, but provides ultimate flexiblity in reaction variations. Standard and HiYield lysates are available with yields of up to 3 mg or up to 5 mg recombinant protein per ml cell-free reaction (Fig.1).

 

Please also refer to our new HiYield-T7 lysates which already contain T7 RNA polymerase, which is required if a T7 promoter driven plasmid is used. For expression regulated by e.g. the tac promoter, addition of this enzyme is not required.

 

Note that our E.coli lysates require additional components (e.g. amino acids, nucleic acids, DNA polymerase, dialysis devices) that are currently not provided by Cube Biotech. Please refer to the protocols for more details.


 

Cell-free protein expression

 

 

Standard vs HiYield Cube E.coli lysates: Protein yields

 

Fig. 1: Standard vs. HiYield
cell-free expression of GFP.
1,7: MW marker, 2-6: HiYield
lysate expression; variation of Mg2+ concentration (14, 16, 18, 20,22 mM). 8-12: Standard lysate expression, same variation of
Mg2+ concentrations. A Mg2+ optimum was observed at ca. 16 mM for these combinations of cell-free lysates and other reaction components. Yields range from 1.3-5 mg/ml for HiYield reactions, and from 1-3.8 mg/ml for standard reactions.
Data kindly provided by Frank Bernhard, University of Frankfurt, Department of Biophysical Chemistry.

 

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E.coli cell lysates

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Cell-free E.coli lysate (350 ul)

Sales price: 99,00 €

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Cell-free E.coli lysate (4x350 ul)

Sales price: 310,00 €

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Cell-free E.coli lysate HiYield (350 ul)

Sales price: 122,00 €

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Cell-free E.coli lysate HiYield (4x350 ul)

Sales price: 386,00 €

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