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Directed evolution, the application of evolutionary design to enzyme engineering, requires effective screening strategies to isolate those proteins that perform a desired function from the libraries generated by the techniques. In Directed Enzyme Evolution: Screening and Selection Methods, seasoned practitioners from many leading laboratories describe their leading and readily reproducible screening strategies for isolating useful clones. These techniques have been optimized for sensitivity, high throughput, and robustness, and are of proven utility for directed evolution purposes. The assays presented use a variety of techniques, including genetic complementation, microtiter plates, solid-phase screens with colorimetric substrates, and flow cytometric screens. There are also representative examples of how phage libraries may be interrogated for enzymatic activity. Each protocol contains detailed step-by-step instructions and many notes on how best to deal with the problems that may occur. An accompanying volume, Directed Evolution Library Creation: Methods and Protocols (ISBN 1-58829-285-1), describes readily reproducible methods for the creation of mutated DNA molecules and DNA libraries. Taken together, Directed Enzyme Evolution: Screening and Selection Methods and Directed Evolution Library Creation: Methods and Protocols capture for newcomers and more experienced investigators alike all the key methods for using directed protein evolution to better understand protein structure-function relationships, to discover new enzymes and therapeutic proteins, and to design new assays suitable for specific applications.

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Since their discovery, Wnt signaling molecules have been shown to control key events in embryogenesis, to maintain tissue homeostasis in the adult and, when aberrantly activated, to promote human degenerative diseases and cancer, thus making them a vital area of study. Wnt Signaling: Methods and Protocols examines both biochemical assays and vertebrate and invertebrate model systems to provide a point of reference to current molecular protocols and the diverse model systems employed to study this important signaling pathway. In Volume 2, Pathway Models, the diverse vertebrate and invertebrate models that have shaped the Wnt signaling field are described, presenting an overview of the unique properties of each organism, like asymmetric cell division in C. elegans and epithelial morphogenesis in Dictyostelium, with respect to studying Wnt/FZD function. As a volume in the highly successful Methods in Molecular Biology™ series, chapters contain readily reproducible laboratory protocols, complete with lists of necessary equipment and reagents and the Notes section, which reveals helpful troubleshooting tips. Comprehensive and cutting-edge, Wnt Signaling: Methods and Protocols collects the expertise and knowledge of many leaders in the field to produce this invaluable two-volume resource. .

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In recent years, a new understanding of cell-penetrating peptides has emerged, helping researchers to expand beyond a number of long-held dogmas. In Cell-Penetrating Peptides: Methods and Protocols, expert researchers explore the latest information on cell-penetrating peptides (CPPs), providing insight into the most important and contemporary areas of CPP research. Chapters address the historical background of CPP studies, provide an overview of the growing field of research, investigate methods for testing CPP mechanisms, present a summary of methods that attempt to use properties of CPPs to study biochemical intracellular mechanisms of interaction and signal transduction, and include new ideas for turning CPP-based strategies into drugs. Composed in the highly successful Methods in Molecular Biology™ series format, each chapter contains a brief introduction, step-by-step methods, a list of necessary materials, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls. Innovative and current, Cell-Penetrating Peptides: Methods and Protocols is an essential guide that allows researchers to study intracellular mechanisms in new ways, and to promote the future development of novel drugs.

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As a high throughput method for analyzing gene function, cell-based microarrays have proven to be of vital importance, allowing high throughput analysis of over expression and knock down of proteins. In Cell-Based Microarrays: Methods and Protocols, experts in the field provide an up to date synopsis of cell-based microarrays and meticulous coverage of all aspects of the array, including emerging technology. Beginning with a detailed overview of the whole subject area, the volume continues with protocols for over-expression arrays and downstream functional assays, infectious disease research, increasing transfection efficiencies, as well as the development of cell-based array technology by use of microfluidic image cytometry for the analysis of small diagnostic samples with few cells. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Comprehensive and cutting-edge, Cell-Based Microarrays: Methods and Protocols serves as a key resource for molecular biologists, geneticists, immunologists, and chemists, and supplies scientists with access to set up a technology that is truly high throughput for the functional analysis of proteins.

Cytology. --- Cell Biology. --- Cell biology --- Cellular biology --- Biology --- Cells --- Cytologists