1-Step Human High-Yield IVT Kit

The Thermo Scientific 1-Step Human High-Yield In Vitro Translation (IVT) Kits enable the expression of functional proteins with 10 to 100 times greater yield per mL than other mammalian IVT systems.

The 1-Step High-Yield IVT System uses modified HeLa cell extracts to take advantage of the robust human translation machinery and generate functional full-length proteins. In this system, protein expression is performed in a proprietary dialysis device that allows a continuous supply of nucleotides, amino acids and energy generating substrates into the reaction while removing inhibitors of proteins synthesis. This continuous-exchange cell-free (CECF) system enables protein expression in an overnight incubation of up to 750 μg/mL. The complete maxi-scale kit includes all the components required for transcription and translation of a recombinant gene, including an optimized expression vector.

Features of the 1-Step Human High-Yield Maxi IVT Kit:

• High expression—up to 750 μg/mL of expressed protein
• Reproducibility—low variability between experiments
• Fast—express high levels of protein with 6 hours to overnight incubation
• Functional—obtain functionally active proteins, including those containing disulfide bonds

1-Step Protein Expression:
• Easy-to-use—transcription and translation is performed in one reaction step
• Adapt for labeling—amenable to incorporation of heavy or unnatural amino acids

Includes:
• Complete kits contain cell lysate, reaction mix, accessory proteins, 5X dialysis buffer, control DNA, cloning vector, dialysis devices and centrifuge tubes

Requires:
• 30°C incubator or water bath (shaker incubator increases expression by 30 to 50%)

Applications:
• Production of recombinant proteins for structural analysis
• Generate active proteins for protein interaction studies
• Perform rapid expression and characterization of mutant proteins
• Small-scale production of viral particles
• Incorporation of unnatural amino acids or toxic protein production

The 1-Step Human High-Yield IVT Kits are cell-free protein expression systems that provide all of the essential components required for transcription and translation. The kits are optimized with Accessory Proteins and Reaction Mixes that support protein synthesis using a DNA template. The advantages of using the 1-Step Human High-Yield IVT Kits over traditional in vivo expression systems include the ability to express toxic or insoluble proteins, easily perform protein labeling with modified amino-acids and reduce the time and cost of expressing human proteins in tissue culture cells. 1-Step Human IVT Kits are optimized for use with the pT7CFE1 expression vector that utilizes the T7 viral promoter and an EMCV Internal Ribosome Entry Site (IRES) to facilitate high levels of in vitro protein expression. Using a pT7CFE vector is critical for obtaining high expression levels. For added convenience, at family of pT7CFE vectors are available with tandem affinity tags for facilitate protein detection and purification. (For smaller scale and screening options, consider the 1-Step Human Coupled IVT Kit.)

More Product Data
• Choosing a vector and purification method for in vitro protein expression
• System for high-throughput, high-yield in vitro translation
• Expression of highly active proteins using a cell-free human IVT system