Next Generation Cell Free Protein Expression Kit (Wheat Germ)

General description

Cell-free protein expression (CFPE) is the production of recombinant proteins in solution using biomolecular translation machinery extracted from cells of different species, such as E. Coli, wheat germ, Leishmania tarentolae, rabbit reticulocyte, insect or human cells. CFPE system from wheat germ extract is suitable for the expression of both prokaryotic and eukaryotic proteins.It usually does not require codon optimization even for the expression of transcripts with a high A/T or G/C content. CFPE system (wheat germ) provides best yield among the eukaryotic systems mentioned above. This system is particularly suitable for production of proteins with complicated conformation and multiple protein complexes, such as disulfide-bond-containing or integral membrane proteins. A variety of proteins and large proteins have been successfully produced from CFPE system (wheat germ), such as human or plant proteins or proteins for vaccine studies. For example, due to its high protein yield and easy to use, CFPE system (wheat germ) was selected as the most suitable choice for a comprehensive protein expression study in the ‘‘Human Protein Factory’’ project at the National Institute of Advanced Industrial Science and Technology (AIST) in Japan. In this study, the researchers tried to express 13364 human proteins, out of which 12996 clones (97.2%) produced protein in CFPE system (wheat germ). They also tested biological activities in two categories of the expressed proteins. Out of the 75 tested phosphatases, 58 (77%) showed biological activity. Also, several cytokines containing disulfide bonds that were expressed in a nonreducing CFPE system (wheat germ) were found in an active form.

Features and Benefits
Next Generation Cell Free Protein Expression Kit (Wheat Germ) (part #: CFPS700) a next generation product protocol to ensure even higher stability, yield and more flexibility for a larger variety of target proteins, such as kinases, membrane proteins transcription factors, antibodies, photoreceptors etc., Our wheat germ extract contains various machines, micelle and organelles. As a result, even membrane protein can be soluble without adding any liposome and disulfide bonds are induced. By using this kit, a protein of interest can be expressed in a day starting from cDNA. There is no need for cloning or cell culture.

• Great Time and cost efficiency, easy to use (batch model)
• Excellent yield: 5.5mL sample scale per kit brings 0.5 mg to 1mg proteins to you in a day at low cost
• No cloning or subcloning necessary for full length or truncation
• Animal- and serum-free
• Of eukaryotic origin
• High solubility due to native conformation and eukaryotic source
• Greater stability
• Suitable for protein labeling and the expression of difficult-to-express proteins like membrane proteins and multiple protein complexes

Components
This product is comprised of the following:

• 1x 25 mM NTP Mix
• 1x T7 RNA Polymerase
• 10x Transcription Buffer
• 1x 100 mM DTT
• 1x Wheat Germ Extract
• 1x Amino Acid Mix

Application

• Expression screening
• High throughput in vitro production of recombinant proteins
• Ribosome Display
• Protein microarrays
• Structural Genomics, NMR (Isotope labeling) or incorporating unnatural amino acid and crystallography

See Instruction for the Primer Design Tool for the 1st PCR
See CFPS700 Primer List (left-click and download the Excel file)

Storage and Stability

Store the components at the designated Temperatures once received.

10x Transcription Buffer: -20 °C
25 mM NTP Mix: -20 °C
T7 RNA Polymerase: -20 °C
100 mM DTT: -20 °C
Wheat Germ Extract: -70 °C
Amino Acid Mix: -70 °C