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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 1, Pathway Methods and Mammalian Models, assays to measure activation of the diverse Wnt signaling pathways are explored, along with a selection of models and strategies used to study mammalian Wnt/FZD function, from protein-protein interaction and simple cell line models to organoid cultures and mouse models. 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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Mitogen-activated protein (MAP) kinase (MAPK) cascades are key signaling components that govern essentially all cellular processes evoked by any type of stimulation, and it has been well established that the malfunctioning of these cascades leads to various diseases including cancer, autoimmunity, and diabetes. In MAP Kinase Signaling Protocols, Second Edition, expert researchers fully update the popular first edition the key techniques used in the study of MAPK signaling cascades in various cellular contexts. This thorough volume explores essential topics such as activation and function of components of the MAPK signaling cascades, the study of MAPK cascades as transmitters of membranal receptor signals, structure-function relationships of MAPKs, studies on the regulation of MAPK cascades, the use of lower organisms, animal models, and human genetics in the study of MAPKs, as well as the study of MAPKs in specific systems and diseases. Written in the highly successful Methods in Molecular Biology™ series format, chapters includes introductions to their respective subjects, lists of the necessary materials, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and cutting-edge, MAP Kinase Signaling Protocols, Second Edition aims to facilitate the study of MAPKs and allow for quicker progress in our knowledge of many vital cellular processes as well as devastating diseases.

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During the past decade, a wide range of scientific disciplines have adopted the use of adipose-derived stem/stromal cells (ASCs) as an important tool for research and discovery. In Adipose-Derived Stem Cells: Methods and Protocols, experts from the field, including members of the esteemed International Federation of Adipose Therapeutics and Science (IFATS), provide defined and established protocols in order to further codify the utilization of these powerful and accessible cells. With chapters organized around approaches spanning the discovery, pre-clinical, and clinical processes, much of the emphasis is placed on human ASC, while additional techniques involving small and large animal species are included. As a volume in the highly successful Methods in Molecular Biology™ series, the detailed contributions 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, Adipose-Derived Stem Cells: Methods and Protocols serves as a vital reference text for experienced researchers as well as new students on the path to further exploring the incredible potential of ASCs.

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