Thermo Scientific 1-Step Human In Vitro
Protein Expression Kits enable the translation and post-transcriptional modification of full-length proteins from mRNA or plasmid templates with yields of up to 100 μg/mL per reaction.
The Human IVT Kits are unique HeLa cell lysate-based protein expression systems for in vitro
translation or coupled transcription/translation reactions. Protein expression is performed in a single 90-minute reaction that can be extended for up to 6 hours with continued protein production up to 100 μg/mL when combined with the optimized pT7CFE1 Expression Vector. The Human IVT Kits can express functional enzymes, phosphoproteins, glycoproteins and membrane proteins for immediate use in studying protein interactions, performing rapid mutational analysis and measuring activity.
Features of the 1-Step Human Coupled IVT Kit:
—uses the human translational machinery to express active proteins
—perform transcription and translation in a single step
• High performance
—greater yields compared to rabbit reticulocyte in vitro
—express proteins that fail in rabbit reticulocyte systemsBetter Than Traditional Methods:
• HeLa cell-free extracts are capable of expressing proteins with post-translational modifications
• Accurate translation delivers full-length protein compatible with downstream applications
• Protein translation is optimized with EMCV IRES element and other mRNA stabilizing elementsIncludes:
• Complete kits contain HeLa cell lysate, accessory proteins, reaction mix, nuclease-free water, expression vector and a GFP-positive control vectorRequires:
• 30°C incubator or water bathApplications:
• Express proteins to measure enzyme activity
• Perform mutational analysis
• Express protein for use in gel mobility shift assays
• Perform co-immunoprecipitation
• Express cytotoxic proteins
• Unnatural amino acid labeling
• Identification and validation of transcriptome small molecule inhibitors
• Screening of RNA targeted small molecules to modify translation
The 1-Step Coupled Human IVT Kit for DNA is a cell-free system using the cellular transcription and translation machinery from a modified HeLa cell line. The procedure is quick and easy and is an effective alternative to other protein expression methods. Simply add an appropriate expression construct to a mixture of HeLa cell lysate, Accessory Proteins, Reaction Mix and incubate at 30°C for 90 minutes to overnight.
The 1-Step Human IVT Kit for mRNA also uses the human protein translation machinery to produce functional protein. This kit is recommended for translation of mRNA transcripts containing an EMCV IRES element and mRNA stabilizing features designed into the pT7CFE1 expression vector.
The human in vitro
translation system is robust and will express proteins from a variety of species including mammals, bacteria and protozoa. Benchmarking shows that the expression levels of functional proteins such as luciferase are much higher in the human system compared to either rabbit reticulocyte-based or E. coli
-based systems. Furthermore, proteins expressed with the human in vitro
translation system are not contaminated with substances that can interfere with downstream applications.More Product Data
• Protocol for in vitro protein expression using mRNA templates
• Protein Biology Application Notes
For Research Use Only. Not for use in diagnostic procedures.