Thermo Scientific™ 1-Step CHO High-Yield IVT Kit

Beschreibung

The Thermo Scientific 1-Step CHO High-Yield In Vitro Translation (IVT) Kit enables the expression of functional proteins with up to 750 times greater yield per mL of reaction than rabbit reticulocyte lysate-based mammalian IVT systems that only yield up to 10μg protein/mL reaction.

• High expression - up to 750μg/mL of expressed protein
• Reproducible results - low variability between experiments
• Fast - express high levels of protein with 6 hours to overnight incubation
• Easier storage - allows you to store lyophilized CHO cell lysate at -20°C
• Functional - obtain functionally active proteins, including those containing disulfide bonds
• Easy-to-use - three reaction components together carry out transcription and translation in one step
• Great for labeling - amenable to incorporation of labeled and unnatural amino acids
• Vector compatibility - uses existing pT7CFE1 vectors for protein expression

The 1-Step CHO High-Yield IVT System uses extracts containing the robust translation machinery of the pharmaceutical industry standard Chinese Hamster Ovary (CHO) cell line to 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 750μg/mL. The CHO IVT kit includes all of the components required for transcription and translation of a recombinant gene, including an optimized expression vector.
Includes:
Lyophilized cell lysate, reaction mix, accessory proteins, 4X 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 large amounts of recombinant protein for structural analysis
• Generation of active proteins for downstream applications such as protein interaction studies
• Rapid large-scale expression and characterization of mutant proteins
• Production of viral particles
• Incorporation of unnatural amino acids
• Production of cytotoxic proteins