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Crystallographic Methods and Protocols by Mark R. Sanderson (auth.), Christopher Jones, Barbara

By Mark R. Sanderson (auth.), Christopher Jones, Barbara Mulloy, Mark R. Sanderson (eds.)

In Crystallographic equipment and Protocols top experts within the program of X-ray crystallography to organic difficulties supply a easy and complete advisor to its winning use in acquiring distinctive three-d buildings of proteins, nucleic acids, and their complexes. those deeply skilled researchers describe in transparent sensible phrases their confirmed equipment for getting ready samples, crystallizing them, acquiring initial characterizations, buying and studying info, and fixing and refining buildings. Their step by step directions provide biologically knowledgeable employees fast entry to a robust strategy necessary to fixing many biologically appropriate difficulties.

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Matthew, C. K. and van Holde, K. E. (1990) Biochemistry, Benjamin-Cummings, Redwood City, CA. Stryer, L. , W. H. Freeman, New York. -I. and Tooze, J. (1991) Chapter 17, in Introduction to Protein Structure, Garland, New York. Cantor, C. R. and Schimmel, P. R. (1980) Part II of Biophysical Chemistry, W. H. Freeman, New York. Eisenberg, D. and Crothers, D. (1979) Chapters 16 and 17, in Physical Chemistry with Applications to the Life Sciences. Benjamin-Cummings Publishing Company, Redwood City, CA.

1987) Generalized method of determining heavy-atom positions using the difference Patterson function. Acta Cryst. A43, 1-5. 10. Sheldrick, G. M. (1990) Phase annealing in SHELX-90: Direct methods for larger structures. Acta Cryst. A46,467--473. 11. , and Woolfson, M. M. (1971) The application of phase relationships to complex structures III. The optimum use of phase relationships. Acta Cryst. A21,410-445. 12. Sheldrick, G. M. , Podjarny, A. , and Thierry, 1. ), Oxford University Press, Oxford, UK.

5-1 % of a 10% solution) or by applica- 34 Skelly and Madden tion to a cellulose-based adsorption media (Cell Debris Remover) supplied by Whatman, Ltd. UK. It is important to note that some loss of protein may occur by coprecipitation. This is especially true of some DNA binding proteins and may be overcome by prior 1: 1 dilution of the crude extract with an appropriate buffer. 2. Primary Isolation Before embarking on the isolation of a novel protein, a specific assay for its activity or a detection method, such as immunoblotting, is an essential prerequisite.

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