in vitro Protein Synthesis Kit PURESYSTEM® Kit flyer download [PDF]
The DnaK set includes purified E.coli DnaK, DnaJ and GrpE.
The GroE set includes purified E.coli GroEL and GroES.
Since DnaK, DnaJ, GrpE and GroES contain a histidine tag at C-terminus, these chaperons can be removed by metal-affinity resin after synthesis. GroEL can also be removed by ultrafiltration as the same method of removing ribosomes in PURESYSTEM.
This product is also applicable for other cell-free protein synthesis and refolding of denatured proteins.
When luciferase and malate dehydrogenase were synthesized in the presence of molecular chaperones, these two proteins were completely soluble and could be easily purified with ultrafiltration and nickel chelating resin.
When luciferase was synthesized in the presence of the DnaK set, enzyme activity of synthesized luciferase was larger than others. In contrast, in the case of malate dehydrogenase (MDH), addition of the GroE set increased the enzyme activity of MDH. Thus, suitable chaperones depend on each synthesizing protein of interest.
Features / Benefits
This product has been designed for helping production of an active form of protein with PURESYSTEM and other technologies. By using this CHAPERONE series with PURESYSTEM is ideal, since other PURESYSTEM components as well as chaperons in this product can be easily removed for purification of protein.