All products have special prices for bulk purchase, please contact for more details if required.

Cat. No.: ECCFS-1 (for the standard version, 1ml)

Cat. No.: ECCFS-10 (for the standard version, 10ml)

Cat. No.: ECCFE-1 (for the enhanced version, 1ml)

Cat. No.: ECCFE-10 (for the enhanced version, 10ml)

Cat. No.: ECCFH-50 (for the high-throughput version, 50ml)

Cat. No.: ECCFH-1k (for the high-throughput version, 1000ml)

Description

The cell-free protein expression system is an in vitro reaction system that utilizes exogenous mRNA or DNA as a template in a reconstituted system containing enzymes and cofactors necessary for cell extracts or protein expression to synthesize proteins. The cell-free protein expression system includes all the templates, molecular machinery, substrates, and energy required for protein expression. It is not limited by the metabolism of the cell itself or the enclosed cellular environment, enabling rapid and high-yield protein expression in vitro. Common cell-free protein expression systems include the Escherichia coli system, insect cell lysate system, wheat germ system, rabbit reticulocyte system, etc.

E. coli Cell-Free Protein Expression Kit operates at the forefront of innovation, leveraging T7 transcription and the innate translation machinery of Escherichia coli. This cutting-edge system encapsulates all vital biological components necessary for transcription, translation, protein folding, and energy regeneration—comprising ribosomes, tRNA, molecular chaperones, metabolic enzymes, translation initiation, and elongation factors. Utilizing PCR products or circular plasmids as templates, the kit streamlines the process, enabling the swift and efficient expression of the target protein. Simply introduce the template, and you're primed for rapid and seamless deployment of your protein synthesis endeavors. Experience the pinnacle of efficiency with our state-of-the-art E. coli Cell-Free Protein Expression Kit.

Principle

The Escherichia coli cell-free protein expression system is currently the most widely used among cell-free protein expression systems. Due to its well-understood genetic background, high expression levels of target genes, short cultivation cycles, high efficiency, low cost, strong resistance to contamination, ease of scale-up, and applicability to the expression of proteins from various species, it is extensively applied in cutting-edge research fields such as protein-protein interactions, high-throughput screening, and gene circuits. Utilizing an efficient transcription and translation coupling reaction, this system produces active recombinant proteins in vitro. The highly efficient and rapid cell-free synthesis system eliminates time-consuming steps in traditional intracellular protein production processes, including transformation, cell cultivation, and expression optimization. Within a few hours, this E. coli cell-free expression system can generate high yields of high-quality proteins. Whether using circular plasmid DNA for protein synthesis, rapid expression screening, or downstream structural or functional analysis, this system significantly simplifies the protein expression process.

Composition

E. coli Cell-Free Protein Expression Kit provides a method for the facile preparation, detection, and purification of recombinant proteins. The expression system offered by this kit primarily includes:

• Optimized Escherichia coli Extracts: These extracts enhance the stability of structures during DNA transcription and translation processes, thereby increasing the yield of soluble proteins.
• Optimized Reaction Buffer: Sustains the provision of energy for protein synthesis through an ATP regeneration system.
• Optimized Concentrations of Amino Acids: Ensures an ample supply of substrates for protein synthesis.
• Optimized Expression Vector Containing GFP Gene: The expression vector includes the GFP gene as a positive control, allowing for the visual observation of protein expression results.

Features

• Rapid: Detection of the target protein can occur in as little as 1 hour, with protein synthesis reaching its peak within 10 hours.
• Template Flexibility: Supports both plasmid and PCR templates, providing versatility in template selection.
• High Expression Levels: Capable of achieving protein yields exceeding 3mg/mL, ensuring high-production protein preparation.
• Versatility in Protein Types: Can express a wide range of protein types, including those rich in disulfide bonds and membrane proteins.
• Simple Operation: A one-step reaction; protein preparation is achieved by simply mixing reaction components with the DNA template.
• High Throughput: Enables expression screening in a 96-well plate format. Upon detecting positive expression, the system can be scaled up.
• Co-expression of Proteins: Allows for the simultaneous expression of multiple proteins in the same reaction.

Transportation and Storage

The kit is shipped with dry ice. After the user opens it, please store the cell-free protein expression reaction components at -80°C for long-term preservation. It is recommended to prepare and use immediately. If it is necessary to divide the reagent after dissolution, it is recommended to aliquot the reagent, flash-freeze with liquid nitrogen, and then store it in a -80°C freezer to avoid repeated freeze-thaw cycles.

Contact us today or drop an email to tech@sbsbio.com for more details and open the door to a world of possibilities!

Related: NTP, Tris Buffered (100mM)

Only for research and not intended for treatment of humans or animals

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