The 1-Step Human Coupled IVT Kit – DNA is a mammalian in vitro translation (IVT) system based on HeLa cell lysates. The kit contains all of the cellular components required for protein synthesis, including ribosomes, initiation factors, elongation factors and tRNA. When supplemented with the included proprietary accessory proteins, reaction mix and a DNA template cloned into the Thermo Scientific pT7CFE1-based vector, this system can synthesise protein for up to 6 hours. The kit includes a pT7CFE-CHis vector for ease of use in downstream applications.
- Uses the human translational machinery to express active proteins
- Perform transcription and translation in a single step
- Greater yields compared to rabbit reticulocyte in vitro translation
- Reliably express proteins that fail in rabbit reticulocyte systems
The benefits of in vitro expression of proteins over traditional in vivo systems include expression of toxic or insoluble proteins, faster protein synthesis and protein labeling with modified amino acids. The optimised kit contains a T7 promoter and an EMCV internal ribosome entry site (IRES) to facilitate high levels of in vitro protein expression in a cap-independent fashion. Using a vector containing the EMCV IRES element is critical for obtaining high expression levels in this human in vitro protein expression system.
Delivery information: Each kit contains HeLa cell lysate (110 or 500 µl), accessory proteins (25 or 100 µl), reaction mix (40 or 200 µl), nuclease-free water (1,5 ml), expression vector (pT7CFE1-CHis - 10 µg) and a GFP-positive control vector (pCFE-GFP - 10 µg).
