Overnight Express™ Autoinduction NMR Medium - ¹5N, ¹3C
Overnight Express™ Autoinduction NMR-15N, -13C, when supplemented with 13C-labeled glycerol (Cat. No. CLM1510EMD), provides reagents to make 1 L of defined autoinducing liquid culture medium used to produce [U-15N, U-13C]-labeled proteins for NMR spectroscopy. The autoinduction method is based on the differential metabolism of carbon sources to promote growth to high cell density and to automatically induce protein expression under control of lac promoters, eliminating the need to monitor cell densities or add inducer (IPTG). The system includes four concentrated sterile solutions: OnEx Dual-NMR Solution 1 is a blend of carbon sources optimized for tightly regulated uninduced growth to high density, followed by induction; OnEx NMR Solution 2 is a buffer and nitrogen blend that mediates metabolic acid production and provides additional nitrogen for increased protein synthesis; OnEx Solution 3 provides magnesium for maximal cell density; OnEx Solution 4 provides trace metals to minimize growth limitations associated with mineral deficiencies and satisfy the requirements of metal-containing target proteins, even at high expression levels. The kit also includes 15N-labeled solid ammonium chloride, 13C-labeled solid glucose, and reagents to make a non-inducing starter medium, for staged growth without expression and for stable storage of clones. Efficient 13C protein labeling requires 13C-glycerol, which must be purchased separately. Addition of the four liquid components and sterile solutions of 15N ammonium chloride, 13C glucose, and 13C glycerol to sterile water results in a chemically- defined medium promoting high cell densities, autoinduction of expression, and efficient labeling of target proteins.
The Overnight Express Autoinduction System is covered by US Patents 7,560,264, 7,704,722, and 7,759,109. This Product is sold for research use only. Any commercial use of the Product, including the manufacture of proteins for use in the screening of compound libraries, requires a commercial license from EMD Millipore Corporation. Commercial use shall include but not be limited to (1) use of the Product or its components in manufacturing; (2) use of the Product or its components to provide a service, information, or data to others in exchange for consideration; (3) use of the Product or its components (or any derivatives thereof) for therapeutic, diagnostic or prophylactic purposes (including as part of a device, chip, assay or other product); or (4) resale of the Product or its components, whether or not such Product or its components are resold for use in research. Nothing contained herein shall be deemed to represent or warrant that additional third party rights are not required for use of this Product.
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