This chapter outlines the procedures and methods required to express, isolate, and purify chemokines in a biologically active form and in high yield in an Escherichia coli expression system. These methods are illustrated with reference to the expression, purification, and characterization of an E. coli-derived recombinant human chemokine known as melanoma growth stimulating activity (MGSA). With this expression system, the recombinant MGSA is secreted into the extracellular medium, which greatly simplifies its isolation. Furthermore, the recombinant chemokine is intact and has the same mass and N-terminal sequence as authentic MGSA. The chapter presents protocols for the analysis and characterization of recombinant MGSA that should greatly aid the investigator embarking on similar projects with other chemokines. The availability of large amounts of chemokines should facilitate progress in the identification of specific receptors for these molecules and help to provide an understanding of their physiological role. Series: Methods in Enzymology (Book 289) Hardcover: 780 pages Publisher: Academic Press; 1 edition (November 4, 1997) Language: English ISBN-10: 0121821900 ISBN-13: 978-0121821906 Product Dimensions: 1.5 x 6.2 x 9.5 inches Link Download http://nitroflare.com/view/E3FE7573DD68E4Fhttps://drive.google.com/drive/folders/1yLBzZ1rSQoNjmWeJTZ3WGQHg04L1
