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The biotechrabbit™ RTS 100 E. coli Disulfide Kit supplies an optimized combination of lysate, chaperones, reagents, and buffer systems to produce maximum yields of active proteins containing multiple disulfide bonds in a CECF system.
The kit employs several innovative technologies, including the optimized high-yield (HY) E. coli lysate, the redox buffer to maintain the system under oxidizing conditions, which enhances the disulfide-bond formation, and disulfide isomerase for rearrangement of disulfide bonds, which improves the yields of correctly folded proteins.
The CECF technology prolongs the reaction time by up to 24 hours, enhancing protein yield. Transcription, translation, and disulfide-bond formation occur in the device's reaction compartment. Substrates and energy components needed for a sustained reaction are continuously supplied via a semipermeable membrane. At the same time, potentially inhibitory interactions with byproducts are diluted by diffusion through the same membrane into the 1 ml feeding chamber. Depending on the particular protein, expression of up to several milligrams of protein per reaction can be obtained (e.g., >2 mg/ml of active urokinase).
