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This four-volume laboratory manual contains comprehensive state-of-the-art protocols essential for research in the life sciences. Techniques are presented in a friendly step-by-step fashion, providing useful tips and potential pitfalls. The important steps and results are beautifully illustrated for further ease of use. This collection enables researchers at all stages of their careers to embark on basic biological problems using a variety of technologies and model systems. This thoroughly updated third edition contains 165 new articles in classical as well as rapidly emerging technologies. Topics covered include: Cell and Tissue Culture: Associated Techniques, Viruses, Antibodies, Immunocyt...
The aim of this book is to provide the researcher with important sample preparation strategies in a wide variety of analyte molecules, specimens, methods, and biological applications requiring mass spectrometric analysis as a detection end-point. In this volume we have compiled the contributions from several laboratories which are employing mass spectrometry for biological analysis. With the latest inventions and introduction of highly sophisticated mass spectrometry equipment sample preparation becomes an extremely important bottleneck of biomedical analysis. We have a goal of giving the reader several successful examples of sample preparation, development and optimization, leading to the success in analytical steps and proper conclusions made at the end of the day. This book is structured as a compilation of contributed chapters ranging from protocols to research articles and reviews. The main philosophy of this volume is that sample preparation methods have to be optimized and validated for every project, for every sample type and for every downstream analytical technique.
Focused manuscript on the potential use/role of miRNAs in bioprocessing, specifically the production of complex proteins in mammalian cells. With that in mind I propose a draft list of topics/chapters along the following lines: Intro on CHO/bioprocessing/engineering challenges to set scene, Genomic organization, biogenesis and mode of action, Identifying miRNA targets: Computational prediction, transcriptomics, proteomices, UTR analysis, etc., miRNA expression in Chinese Hamster Ovary cells, miRNAs as engineering targets: pathway manipulation to impact bioprocess phenotypes, miRNAs as biomarkers, Detection methods: Northern, PCR, hybridization arrays, Next Gen Seq, Manipulation of expression in cultured cells: Transient/stable disregulation, Knockout.
Cell culture techniques allow a variety of molecular and cell biological questions to be addressed, offering physiological conditions whilst avoiding the use of laboratory animals. In addition to basic techniques, a wide range of specialised practical protocols covering the following areas are included: cell proliferation and death, in-vitro models for cell differentiation, in-vitro models for toxicology and pharmacology, industrial application of animal cell culture, genetic manipulation and analysis of human and animal cells in culture.
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The second edition of this volume provides a comprehensive update of this key method on gel-based proteomics. Chapters present an introduction into the development of methods on principles of differential protein labeling and two-dimensional gel electrophoresis, techniques on optimized proteomic workflows using advanced mass spectrometry for protein identification, and the application of those methods in basic biological research, pathobiology and applied biomarker discovery. 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. Authoritative and cutting-edge, Difference Gel Electrophoresis: Methods and Protocols, Second Edition aims to ensure successful results in the further study of this vital field.
This third edition expands on the previous editions with updated and new chapters on protein chromatography. Chapters detail protein stability and storage, avoiding proteolysis, protein quantitation methods, generation and purification of recombinant proteins, recombinant antibody production, and the tagging of proteins. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Protein Chromatography: Methods and Protocols, Third Edition aims to provide commonly used methods and new approaches to help both new researchers and experts expand their knowledge.
This detailed new edition explores the use of Chinese hamster ovary (CHO) cells in the production of therapeutic protein products. Beyond updates on earlier methodologies, the book also delves into the genetic manipulation of CHO cells for recombinant protein production, analysis of CHO cells using proteomic and metabolomic approaches, as well as methods for the characterization of recombinant protein products, such as glycosylation and host cell protein analysis. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Heterologous Protein Production in CHO Cells: Methods and Protocols, Second Edition is an ideal guide for researchers working to enhance and accelerate CHO productive capabilities in the coming decades.
Proteomics Mass Spectrometry Methods: Sample Preparation, Protein Digestion, and Research Protocols shares best practices collected across key laboratories and core facilities, taking the reader through key tactics for executing the most usual mass spectrometry experiments. Sections review research making use of MS proteomics experiments, focus on critical sample preparation, cover mammalian cell lines and samples from clinical tissue and biological fluids, discuss subcellular fractionation, provide methods for protein digestion both for in gel and in solution, and delve into key MS proteomics analysis protocols, including label-free LC-MS, TMT and iTRAQ labelled LC-MS, phosphorylation enric...