Subject Search: Protein Expression

Glycoprotein expression in a human IVT system

Glycosylation is a form of co-translational and post-translational modification in which sugars are added onto proteins. Glycosylation is important for protein function, protein folding and cell-cell adhesions. The Thermo Scientific 1-Step Human Coupled IVT Kit (Part No. 88881) contains HeLa cell lysate and other components to supply the cellular machinery required to support protein translation

Continue reading…

Expression of highly active proteins using a cell-free human IVT system

In vitro translation (IVT) systems have been used for rapid expression of functional proteins, including enzymes. Typically, proteins expressed in mammalian cells have higher activity compared to those expressed in bacteria, because slower translational rates in the mammalian systems indirectly influence better folding of proteins. The Thermo Scientific 1-Step Human High-Yield IVT System is based

Continue reading…

System for high-throughput, high-yield in vitro translation

Cell-free, in vitro translation (IVT) systems enable small amounts of proteins to be produced quickly. Protein expression protocols that can be adapted to high-throughput (HTP) techniques provide an efficient means for producing a large number of recombinant proteins for use in screening and other research applications. Here we demonstrate the speedy and efficient simultaneous expression

Continue reading…

Choosing a vector and purification method for in vitro protein expression

In vitro protein expression is a rapid technique to produce functional proteins. By contrast with in vivo protein expression methods, in vitro protein expression allows for expression of toxic proteins, incorporation of heavy isotopes for NMR or MS analysis, addition of non-natural amino acids, and elimination of growth and induction conditions for recombinant protein synthesis.

Continue reading…

Protocol for in vitro protein expression using mRNA templates

The Thermo Scientific 1-Step Human Coupled IVT Kit (Part No. 88881) is a mammalian in vitro translation (IVT) system based on HeLa cell lysates for use with DNA templates. The kit contains all of the cellular components required for protein synthesis, including ribosomes, initiation factors, elongation factors and tRNA. Transcription and translation occur in a

Continue reading…

Simultaneous dual-emission detection of luciferase reporter assays

Luciferase-based reporter assays are a powerful tool for monitoring gene expression in cells because of their ultrasensitive detection capacity and wide dynamic range. These assays involve inserting a genetic regulatory element upstream of a luciferase gene and then transferring the resulting reporter construct into animal cells, plant cells or bacteria via transfection, transformation or injection.

Continue reading…

Luciferase Detection Instrumentation Guide

Luciferase reporter assays are widely used because of their sensitive detection capacity and broad dynamic range. And while many companies make instruments to detect and measure luciferase activity, identifying the right instrument for you can be difficult because of their high feature variability. We therefore gathered salient information about many of the instruments available to

Continue reading…

Secreted dual-spectral luciferase reporter assays

The study of complex cellular signaling pathways requires powerful and specific tools to monitor changes in gene activation or repression. To accurately monitor these processes, reporter gene assays are commonly used. Our multiplexed luciferase reporters for studying gene regulation are more sensitive and convenient than luciferase reporter systems from other suppliers. We use two naturally

Continue reading…