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CFPS700 Sigma-Aldrich

Next Generation Cell Free Protein Expression Kit (Wheat Germ)

An Eukaryotic Cell Free Protein Expression system (CFPE/CFPS) with High Yield and Excellent Scalability, 50 reactions

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Properties

shipped in   dry ice

Description

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.

See for most Frequently Asked Questions here.

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
See datasheet for more information.

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

Safety & Documentation

Safety Information

Safety Information for this product is unavailable at this time.

Documents

Certificate of Analysis

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Protocols & Articles

Articles

FAQs for Next Generation Cell Free Protein Expression Kit (Wheat Germ) - CFPS700

Easily scalable protein production using Next Generation Cell Free Protein Expression Kit (Wheat Germ).
Keywords: Dialysis, Electrophoresis, Gene expression, Phosphorylations, Polymerase chain reaction, Post translational modifications, Transcription, transformation

Introduction to Cell-Free Protein Expression/Synthesis (CFPE/CFPS)

A form of synthetic biology, cell-free protein expression (CFPE) is a rapid method to produce recombinant proteins in solution without involving cells. Using biomolecular translation machinery extrac...
Keywords: Cell culture, Cell disruption, Cloning, Degradations, Gene expression, Genetic, Microarray Analysis, Polymerase chain reaction, Protein microarrays, Research areas, Transcription, Western blot

Primer Design Tool for the 1st PCR and Instruction of how to use this tool

The 2-step PCR is the first step in the protocol of CFPS700 kit to generate the DNA template to be used in the following transcription and translation. The 1st PCR aims to connect the target ORF sequ...
Keywords: Gas chromatography, PAGE, Polymerase chain reaction, Transcription

Protocols

Next Generation Cell Free Protein Expression Kit (Wheat Germ) (CFPS700) PROTOCOL

Protein synthesis using cell-free protein synthesis reagent kit consists of three stages: preparation of transcription template, transcription and translation. For the use of the translation reagent ...
Keywords: AGE, Amplification, Dialysis, Electrophoresis, Evaporation, Gel electrophoresis, Gene expression, Immunostaining, Polymerase chain reaction, Purification, Size-exclusion chromatography, Transcription, Western blot

Related Content

Quick Video Tutorials for Next Generation Cell Free Protein Expression Kit (Wheat Germ) – CFPS700

The following videos provide quick tutorials on the three major steps following PCR when using the Next Generation Cell Free Protein Expression Kit (Wheat Germ) (CFPS700): transcription, mRNA purific...
Keywords: Gene expression, Polymerase chain reaction, Purification, Transcription

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