Production of human leukocyte extract and Nluc expression. (A) Leukopak was purchased form the AllCells, Inc. (B) The cells were clarified by centrifugation followed by washing with wash buffer (see materials and methods section). A white leukocyte layer is seen in the top layer of the centrifuged sample. (C) Washed cells were reconstituted in extraction buffer and lysed using nitrogen cavitation method (D) Leukocyte extract collected from the vessel was further clarified and the final extract was collected in Eppendorf tubes and stored at −80°C. (E) Expression of Nano-luciferase in leukocytes extracts - the Leukocyte cell-free protein synthesis system consists of leukocyte cell lysate and solutions for reaction mix and dialysis buffer. In vitro transcription and translation reactions (IVT) were setup in micro dialysis device for overnight expression (~16 h) and samples were collected and tested for luciferase activity.