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Over the past thirty years, the development of the Western blot has revolutionized the fields of biomedical research and medical diagnostics. In "Protein Blotting and Detection: Methods and Protocols", expert researchers present numerous techniques based on the Western blot, providing detailed, readily reproducible methods, tips, and alternatives directly and easily transferable to the laboratory setting. Chapters offer a large number of variations on the theme of protein transfer to solid support followed by detection, presenting both adaptations of traditional techniques as well as completely original methods of protein blotting. Composed in the highly successful Methods in Molecular BiologyTM series format, each chapter contains a brief introduction, a list of necessary materials, step-by-step methods, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls. Innovative and highly practical, Protein Blotting and Detection: Methods and Protocols is an essential, hands-on guide for all investigators who hope to bring these cutting-edge procedures home to their laboratories.
Over the past thirty years, the development of the Western blot has revolutionized the fields of biomedical research and medical diagnostics. In "Protein Blotting and Detection: Methods and Protocols", expert researchers present numerous techniques based on the Western blot, providing detailed, readily reproducible methods, tips, and alternatives directly and easily transferable to the laboratory setting. Chapters offer a large number of variations on the theme of protein transfer to solid support followed by detection, presenting both adaptations of traditional techniques as well as completely original methods of protein blotting. Composed in the highly successful Methods in Molecular BiologyTM series format, each chapter contains a brief introduction, a list of necessary materials, step-by-step methods, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls. Innovative and highly practical, Protein Blotting and Detection: Methods and Protocols is an essential, hands-on guide for all investigators who hope to bring these cutting-edge procedures home to their laboratories.
This book fills the need for a simplified text covering western blotting protocols aimed not just at high school and college students, but the researcher with little to no experience in these techniques. It provides the principles, basic methodology, and tips and tricks to avoiding the common pitfalls of western blotting. The book also introduces simple protocols that can transform western blotting into a fun method, such as sending secret messages on membranes or using nitrocellulose membrane as a canvas for art. In addition to the techniques, this book also covers the history of western blotting, which originated from the development of the blotting of DNA. It then delves into the importance of protein blotting, brought to the fore by the fact that the procedure has been evolving constantly since its inception in 1979, and the fact that the scientific community is faced with a multitude of ways and means of transferring proteins to membranes..
Western Blotting Guru provides researchers in molecular biology with a handy reference for approaching and solving challenging problems associated with immunoblotting setup and optimization. As a laboratory guide, it emphasizes the technical aspects of efficiently employing immunoblotting as a tool in molecular biology laboratories. The book covers the basic science underlying immunoblotting and detailed description of the method parameters, followed by good benchtop practices, tips and tricks for obtaining high-quality data and a detailed troubleshooting guide addressing a variety of problem types. - Provides a benchtop reference that every molecular biologists will use to design, optimize, troubleshoot and analyze their immunoblotting experiments - Contains unique good practices and tips that are indispensable for the beginner and expert alike - Features special cases with applications of immunoblotting optimization - Includes detailed appendices with tables, figures and key protocols - Provides troubleshooting tips for various types of modifications of standard protocols - Organized as a systematic, concentrated resource to save time when addressing an immunoblotting problem
The authors are commonly the techniques" originators, and each has demonstrated a hands-on mastery of the methods described, always fine-tuning them here for optimal productivity.
This volume covers past and present western blot techniques, such as diffusion blotting, slice blotting, blotting of high and low molecular weight proteins, single cell blotting and automated blotting. 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 tips on troubleshooting and avoiding known pitfalls. Thorough and cutting-edge, Western Blotting: Methods and Protocols will serve as an invaluable reference for those interested in further study into this fascinating field.
This laboratory manual includes the latest tools and techniques involved in genomic research. It starts with an introductory chapter on genomics and the various tools and applications involved. The initial chapters present protocols for basic techniques such as DNA isolation, electrophoresis, PCR, cDNA synthesis etc. The book then goes on to describe more advanced techniques such as next-generation sequencing, exome sequencing, use of RNAi, RNAseq, genome editing, single cell genomics etc. Each topic includes a brief description, information on the principles involved, materials & methods, protocol, and expected results, with diagrams and graphs. All protocols are presented in a very lucid and precise way, to make it easy for readers to follow and replicate them.
T. T. Ngo and H. M. Lenhoff Department of Developmental and Cell Biology University of California, Irvine, CA 92717 In 1959, Yalow and Berson used insulin labeled with radioactive iodine to develop a quantitative immunological method for determining the amount of insulin in human plasma. Their method depends upon ~ competition between insulin labeled with radioactive iodine (II 1) and unlabeled insulin from plasma for a fixed and limited number of specific binding sites on the antibody to insulin. The amount of the labeled insulin bound to the antibody is inversely proportional to the amount of insulin in the plasma sample. Their method, which is so elegantly simple in concept, is made possible by the ability to detect with ease extremely low levels of radioactivity, and by the exquisite specificity of an antibody capable of specifically binding the analyte. Such a combination of sensitivity and specificity is the basis of this versatile analytical tool called radioimmunoassay (RIA). Twelve years later, Engvall and Perlmann (1971) and Van Weemen and Schuurs (1971) independently introduced the use of enzymes as another category of sensitive and even more versatile labels for use in immunoassays. Engvall and Perlmann (l971) coined the term ELISA, which stands for Enzyme Linked Immunosorbent Assay.
This work provides rapid access to focused information on topics of Immunotoxicology not only for scientists and those dealing with laboratory aspects but also for lecturers and advanced students. Over 200 contributing authors – including many of the world’s top specialists – have contributed full essays on all relevant topics, supplemented by keyword definitions of related terms. Full essays are structured uniformly to provide reader-friendly information on all aspects of Immunotoxicology, including methods of testing and analysis, characteristics of substances, the regulatory environment and the relevance of these to humans. The single A–Z format of both types of entry makes this reference book very easy to use. The Encyclopedic Reference of Immunotoxicology is intended to be a comprehensive work of reference which will provide easy access to relevant information in the fast-growing field of Immunotoxicology.
Polymers and Nanomaterials for Gene Therapy provides the latest information on gene therapy, a topic that has attracted significant attention over the past two decades for the treatment of inherited and acquired genetic diseases. Major research efforts are currently focused on designing suitable carrier vectors that compact and protect oligonucleotides for gene therapy. The book explores the most recent developments in the field of polymer science and nanotechnology, and how these advancements have helped in the design of advanced materials. Non-viral vector systems, including cationic lipids, polymers, dendrimers, peptides and nanoparticles, are potential routes for compacting DNA for systemic delivery. However, unlike viral analogues that have no difficulty in overcoming cellular barriers and immune defense mechanisms, non-viral gene carriers consistently exhibit significant reduced transfection efficiency due to numerous extra- and intracellular obstacles. Therefore, biocompatibility and potential for large-scale production make these compounds increasingly attractive for gene therapy. This book contains chapters on the engineering of polymers and nanomaterials for gene therapy, and how they can form complexes with DNA and avoid both in vitro and in vivo barriers. Other chapters describe in vitro, ex vivo, in vivo gene therapy studies, and the current issues affecting non-viral gene therapy. - Explores current challenges in the research of genetic diseases - Discusses polymers for gene therapy and their function in designing advanced materials - Provides examples of organic and inorganic nanomaterials for gene therapy - Includes labeling, targeting, and assays - Looks at characterization, physico-(bio)chemical properties, and applications