Product Overview :
E. coli cell-free protein expression method is the coupled reaction of transcription and translation. The method use a mixture containing template DNA bearing a gene of interest (either expression vector or PCR product), E. coli cell extract and other required reagents such as amino acids and rNTPs. rNTPs, and T7 RNA polymerase synthesize mRNA from template DNA, and then ribosome and tRNA are required for the translation step.
Ni-NTA magnetic beads have many advantages over existing resins and are suited for purification of His-tagged recombinant protein. Ni-NTA groups coated on surface of the beads can interact with His-tag of expressed protein to make that protein to bind them on the bead. After washing out unbound proteins, the target proteins can be purified through elution process.
Description :
Protein Synthesis Kit uses a manual procedure for protein expression and purification. It includes all components required for cell-free protein expression using E. coli extract and Ni-NTA affinity purification. Target protein is recovered in about 100 μg per reaction.
Kit Components :
This kit consists of two boxes (Kit① and Kit②). The Kit① and Kit② contains components for Ni-affinity purification and
for E. coli cell-free protein expression, respectively.
Kit① (store at 4 ~ 8°C):
Ni-NTA magnetic bead: 50 mg x 5 tubes;
Elution buffer: 1.25 mL x 1 tube;
Binding/washing buffer: 25 mL x 1 bottle.
Kit② (store at -20°C ~ - 70°C):
E. coli extract： 200 uL × 5 tubes;
Master mix： 350 uL × 5 tubes;
DEPC DW： 1.0 mL × 1 tube;
Positive Control DNA： 10 µL x 1 tube.
Features & Benefits :
High-speed: synthesize your target protein quickly (within 3 hours) and economically
Easy-to-Use: includes all necessary components for transcription and translation
Flexible: synthesize proteins from various DNA templates.
synthesize proteins of toxic proteins
incorporate non-natural amino acids
use additives such as metal ions, detergents, cofactors, binding partners
PDF