RTS 100 E. coli Disulfide Kit

The RTS 100 E. coli Disulfide Kit provides an optimized combination of lysate, reagents, chaperones, and buffer systems to produce maximum yields of active multiple-disulfide-bonded protein in a CECF (continuous-exchange, cell-free) system.
The kit employs several innovative technologies:

• Coupled transcription/translation system; no mRNA isolation necessary
• Optimized high-yield (HY) E. coli lysate
• Redox buffer to maintain the system under oxidizing conditions, which enhances disulfide-bond formation
• Disulfideisomerase for rearrangement of disulfide bonds, which enhances yields of correctly folded proteins
• Increased amounts of GroE chaperones, which produce higher yields of soluble, properly folded proteins

It utilizes CECF-technology, which prolongs the reaction time for at least 24 hours and therefore enhances the yield of produced proteins. Transcription, translation, and disulfide-bond formation take place in the 50µl reaction compartment of the reaction device. Substrates and energy components needed for a sustained reaction are continuously supplied via a semipermeable membrane. At the same time, potentially inhibitory reaction by-products are diluted via diffusion through the same membrane into the 1 ml feeding chamber. Depending on the particular protein, expression of up to 50 µg protein per 50 µl reaction can be obtained.