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.

Describes and integrates the techniques of many advances in both chromatographic and mass spectrometric technologies. This book also covers various biophysical applications, such as H/D exchange for study of conformations, protein-protein and protein-metal and ligand interactions. It also describes atto-to-zepto-mole quantitation of 14C and 3H.

Introduces readers to the state of the art of omics platforms and all aspects of omics approaches for clinical applications This book presents different high throughput omics platforms used to analyze tissue, plasma, and urine. The reader is introduced to state of the art analytical approaches (sample preparation and instrumentation) related to proteomics, peptidomics, transcriptomics, and metabolomics. In addition, the book highlights innovative approaches using bioinformatics, urine miRNAs, and MALDI tissue imaging in the context of clinical applications. Particular emphasis is put on integration of data generated from these different platforms in order to uncover the molecular landscape of diseases. The relevance of each approach to the clinical setting is explained and future applications for patient monitoring or treatment are discussed. Integration of omics Approaches and Systems Biology for Clinical Applications presents an overview of state of the art omics techniques. These methods are employed in order to obtain the comprehensive molecular profile of biological specimens. In addition, computational tools are used for organizing and integrating these multi-source data towards developing molecular models that reflect the pathophysiology of diseases. Investigation of chronic kidney disease (CKD) and bladder cancer are used as test cases. These represent multi-factorial, highly heterogeneous diseases, and are among the most significant health issues in developed countries with a rapidly aging population. The book presents novel insights on CKD and bladder cancer obtained by omics data integration as an example of the application of systems biology in the clinical setting. Describes a range of state of the art omics analytical platforms Covers all aspects of the systems biology approach—from sample preparation to data integration and bioinformatics analysis Contains specific examples of omics methods applied in the investigation of human diseases (Chronic Kidney Disease, Bladder Cancer) Integration of omics Approaches and Systems Biology for Clinical Applications will appeal to a wide spectrum of scientists including biologists, biotechnologists, biochemists, biophysicists, and bioinformaticians working on the different molecular platforms. It is also an excellent text for students interested in these fields.

With usage of mass spectrometry continually expanding, an increasing number of scientists, technicians, students, and physicians are coming into contact with this valuable technique. Mass spectrometry has many uses, both qualitative and quantitative, from analyzing simple gases to environmental contaminants, pharmaceuticals, and complex biopolymers

The phenotype of a plant in response to a stress condition is the reflection of the molecular responses in different cell-types composing the plant. The multicellular complexity represents a challenge when accessing specific responses of each cell or cell type composing the plant. To overcome this difficulty and allow the clear characterization of the plant cell molecular mechanisms, the research community is now focusing on studying a single cell and single cell-types. The isolation of plant single cells is limited by the cell wall that confers the rigidity of the plant and its overall structure. Various methods have been developed for isolating plant cells (e.g. laser capture microdissection; cell sorting of Green Fluorescent Protein (GFP)-tagged protoplasts, differential protoplastization of cells such as guard cells, isolation of easily accessible cell types such as cotton fiber, pollen cells, trichomes and root hair cells). The development of these innovative approaches to isolate single plant cells or cell-types combined with the application of sensitive and high-throughput technologies allows a better analysis of the developmental processes and response to environmental stresses. Ultimately, single plant cell and cell-type biology will lead to establishment of more reliable and accurate -molecular regulatory networks at the resolution of basic life unit. The goal of this Research Topic is to cover new technological and biological advances in the study of plant single cell, cell-type and systems biology.

This long-awaited first guide to sample preparation for proteomics studies overcomes a major bottleneck in this fast growing technique within the molecular life sciences. By addressing the topic from three different angles -- sample, method and aim of the study -- this practical reference has something for every proteomics researcher. Following an introduction to the field, the book looks at sample preparation for specific techniques and applications and finishes with a section on the preparation of sample types. For each method described, a summary of the pros and cons is given, as well as step-by-step protocols adaptable to any specific proteome analysis task.

Provides a comprehensive description of mass spectrometry basics, applications, and perspectives Mass spectrometry is a modern analytical technique, allowing for fast and ultrasensitive detection and identification of chemical species. It can serve for analysis of narcotics, counterfeit medicines, components of explosives, but also in clinical chemistry, forensic research and anti-doping analysis, for identification of clinically relevant molecules as biomarkers of various diseases. This book describes everything readers need to know about mass spectrometry—from the instrumentation to the theory and applications. It looks at all aspects of mass spectrometry, including inorganic, organic, forensic, and biological MS (paying special attention to various methodologies and data interpretation). It also contains a list of key terms for easier and faster understanding of the material by newcomers to the subject and test questions to assist lecturers. Knowing how crucial it is for young researchers to fully understand both the power of mass spectrometry and the importance of other complementary methodologies, Mass Spectrometry: An Applied Approach teaches that it should be used in conjunction with other techniques such as NMR, pharmacological tests, structural identification, molecular biology, in order to reveal the true function(s) of the identified molecule. Provides a description of mass spectrometry basics, applications and perspectives of the technique Oriented to a broad audience with limited or basic knowledge in mass spectrometry instrumentation, theory, and its applications in order to enhance their competence in this field Covers all aspects of mass spectrometry, including inorganic, organic, forensic, and biological MS with special attention to application of various methodologies and data interpretation Includes a list of key terms, and test questions, for easier and faster understanding of the material Mass Spectrometry: An Applied Approach is highly recommended for advanced students, young scientists, and anyone involved in a field that utilizes the technique.

Two new volumes of Methods in Enzymology continue the legacy of this premier serial with quality chapters authored by leaders in the field. Circadian Rhythms and Biological Clocks Part A and Part B is an exceptional resource for anybody interested in the general area of circadian rhythms. As key elements of timekeeping are conserved in organisms across the phylogenetic tree, and our understanding of circadian biology has benefited tremendously from work done in many species, the volume provides a wide range of assays for different biological systems. Protocols are provided to assess clock function, entrainment of the clock to stimuli such as light and food, and output rhythms of behavior and physiology. This volume also delves into the impact of circadian disruption on human health. Contributions are from leaders in the field who have made major discoveries using the methods presented here. - Continues the legacy of this premier serial with quality chapters authored by leaders in the field - Covers research methods in biomineralization science - Keeping with the interdisciplinary nature of the circadian rhythm field, the volume includes diverse approaches towards the study of rhythms, from assays of biochemical reactions in unicellular organisms to monitoring of behavior in humans.

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.

Zusammenfassung: This edited volume discusses the identification, discovery, characterization, structure determination and modeling of multicomponent macromolecular complexes, and as such, it fully complements the first volume (ISBN 978-3-319-27214-6), which targeted methods of recombinant production of protein complexes. This book is divided in 8 sections offering a selection of technologies widely used in the characterization of protein and protein-nucleic acid complexes for different purposes and at different scales. From native electrophoresis methods, that are accessible to any reasonably well-equipped laboratory, to the sophisticated setup required for structure determination by cryo-electron microscopy or X-ray crystallography, this book contains a wide variety of clearly explained analytic and preparative approaches, connected with the production techniques developed on the previous volume. The readers will find an integral connection between this book and the first volume, which ensures a comprehensive and updated discussion of the main topics of the discipline. Taken together, these volumes constitute a cohesive and authoritative source of the research on multicomponent macromolecular complexes. In here, we focus on characterization of protein complexes in the broadest sense, which is not typically covered in other sources. Moreover, all chapters are carefully written by world renowned scientists and active researchers, making this volume ideal, not only as a reference source, but also as a companion book for the daily laboratory work. This book is aimed for a wide range of scientists, from science students to experienced researchers, working on protein and protein-nucleic acid complexes, who need a thorough understanding of protein production and complex characterization.