In vitro translation (IVT) using mammalian cell extracts is a quick and convenient alternative to in vivo mammalian protein expression. Thermo Fisher has developed IVT systems from two mammalian cell lines, HeLa and CHO, which enable rapid expression of genes from mammals, bacteria or protozoa. The Thermo Scientific 1-Step Human Coupled, 1-Step Human High-Yield, and 1-Step CHO High-Yield IVT kits allow rapid protein expression through the addition of a DNA of interest to a mixture of HeLa or CHO cell lysate and reaction mix followed by incubation for 1-6 hours at 30°C. For optimal expression in both IVT systems a series of vectors (pT7CFE1) with a variety of purification tags including Gateway™ compatible vectors for high throughput applications have been engineered. Thermo Fisher will present data demonstrating expression, purification, and activity of several different proteins using both the HeLa and CHO IVT kits. Results indicate that highly functional proteins can be produced in milligram quantities for a variety of downstream applications. Further, a new protein service option will be discussed encompassing gene synthesis, IVT expression, and purification with turnaround time less than 10 business days.