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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.
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
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..
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 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.
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 is a practical biochemical guide to the Enzyme-Linked Immunosorbent Assay (ELISA), used to detect a target substance in a liquid sample. The ELISA is an important and widely used diagnostic tool in medicine, animal health, botany and quality assurance processes in food and beverage production. An introductory chapter orients the reader on the basic structure and function of immunoglobulins and their fragments while subsequent chapters outline the methodology to generate monoclonal antibodies using hybridoma technology and the general methods used to purify antibodies. Multiple chapters demonstrate how to creatively use the properties of the antibody to identify, localize and quantify target analytes to answer questions and resolve problems. The reader will learn how to use a variety of immunoassay strategies, reporters and detection systems that will undoubtedly facilitate their efforts to gain answers to their own questions. Written in the 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 protocols and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, ELISA: Methods and Protocols seeks to provide both professionals and novices with the technical information necessary for the reader to successfully use the immunoassay as part of the discovery process.
Basic Science Methods for Clinical Researchers addresses the specific challenges faced by clinicians without a conventional science background. The aim of the book is to introduce the reader to core experimental methods commonly used to answer questions in basic science research and to outline their relative strengths and limitations in generating conclusive data. This book will be a vital companion for clinicians undertaking laboratory-based science. It will support clinicians in the pursuit of their academic interests and in making an original contribution to their chosen field. In doing so, it will facilitate the development of tomorrow's clinician scientists and future leaders in discovery science. - Serves as a helpful guide for clinical researchers who lack a conventional science background - Organized around research themes pertaining to key biological molecules, from genes, to proteins, cells, and model organisms - Features protocols, techniques for troubleshooting common problems, and an explanation of the advantages and limitations of a technique in generating conclusive data - Appendices provide resources for practical research methodology, including legal frameworks for using stem cells and animals in the laboratory, ethical considerations, and good laboratory practice (GLP)
This manual is designed as an intensive introduction to the various tools of molecular biology. It introduces all the basic methods of molecular biology including cloning, PCR, Southern (DNA) blotting, Northern (RNA) blotting, Western blotting, DNA sequencing, oligo-directed mutagenesis, and protein expression. - Provides well-tested experimental protocols for each technique - Lists the reagents and preparation of each experiment separately - Contains a complete schedule of experiments and the preparation required - Includes study questions at the end of each chapter
This volume explores the use of mass spectrometry for biomedical applications. Chapters focus on specific therapeutic areas such as oncology, infectious disease and psychiatry. Additional chapters focus on methodology as well as new technologies and instrumentation. This volume provides readers with a comprehensive and informative manual that will allow them to appreciate mass spectrometry and proteomic research but also to initiate and improve their own work. Thus the book acts as a technical guide but also a conceptual guide to the newest information in this exciting field. Mass spectrometry is the central tool used in proteomic research today and is rapidly becoming indispensable to the biomedical scientist. With the completion of the human genome project and the genomic revolution, the proteomic revolution has followed closely behind. Understanding the human proteome has become critical to basic and clinical biomedical research and holds the promise of providing comprehensive understanding of human physiological processes. In addition, proteomics and mass spectrometry are bringing unprecedented biomarker discovery and are helping to personalize medicine.