Basic Principles Of Analytical Ultracentrifugation

Analytical ultracentrifugation (AUC) can supply rich information on the mass, shape, size distribution, solvation, and composition of macromolecules and nanoscopic particles. It also provides a detailed view of their reversible single- or multi-component interactions over a wide range of affinities. Yet this powerful technique has been hard to master in mainstream molecular sciences due to a lack of comprehensive books on the subject. Filling this gap in the literature of biophysical methodology, Basic Principles of Analytical Ultracentrifugation explains the fundamentals in the theory and practice of AUC. The book provides you with up-to-date experimental information to confidently practice AUC. You will understand the basic concepts, full potential, and possible pitfalls of AUC as well as appreciate the current relevance of past work in the field. The book first introduces the basic principles and technical setup of an AUC experiment and briefly describes the optical systems used for detection. It then explores the ultracentrifugation experiment from a macromolecular standpoint, offering a detailed physical picture of the sedimentation process and relevant macromolecular parameters. The authors present important practical aspects for conducting an experiment, including sample preparation, data acquisition and data structure, and the execution of the centrifugal experiment. They also cover instrument calibration and quality control experiments.

Analytical ultracentrifugation is one of the most powerful solution techniques for the study of macromolecular interactions, to define the number and stoichiometry of complexes formed, and to measure affinities ranging from very strong to very weak and repulsive. Building on the data analysis tools described in the volume Sedimentation Velocity Analytical Ultracentrifugation: Discrete Species and Size-Distributions of Macromolecules and Particles, and the experimental and instrumental aspects in the first volume Basic Principles of Analytical Ultracentrifugation, the present volume Sedimentation Velocity Analytical Ultracentrifugation: Interacting Systems is devoted to the theory and practical data analysis of dynamically coupled sedimentation processes. This volume is designed to fill a gap in biophysical methodology to provide a framework that builds on the fundamentals of the highly developed traditional methods of analytical ultracentrifugation, updated with current methodology and from a viewpoint of modern applications. It will be an invaluable resource for researchers and graduate students interested in the application of analytical ultracentrifugation in the study of interacting systems, such as biological macromolecules, multi-protein complexes, polymers, or nanoparticles.

A follow-up to the experimental and instrumental aspects described in Basic Principles of Analytical Ultracentrifugation, the volume Sedimentation Velocity Analytical Ultracentrifugation: Discrete Species and Size-Distributions of Macromolecules and Particles describes the theory and practice of data analysis. Mathematical models for the sedimentation process and the evolution of detected signals are developed in a comprehensive framework, jointly with the description of current and historical strategies for how to extract from noisy experimental data the physical parameters of interest, such as size, mass, and shape, composition, and polydispersity of sedimenting particles. The methods are extensively illustrated, and supported with practical applications, as well as cross-references where to find the methods in the public domain software SEDFIT and SEDPHAT. The systems covered are discrete or polydisperse mixtures of sedimenting molecules or particles in dilute solution, such as proteins and other biomolecules and their stable complexes, man-made polymers, and nanoparticles, observed in different optical systems. A useful reference for researchers and graduate students of macromolecular disciplines, these methods form the essential foundation for the analysis of dynamic interacting systems, which are covered in the volume Sedimentation Velocity Analytical Ultracentrifugation: Interacting Systems. Software referenced in the book is available for download at: https://sedfitsedphat.nibib.nih.gov/default.aspx

A follow-up to the experimental and instrumental aspects described in Basic Principles of Analytical Ultracentrifugation, the volume Sedimentation Velocity Analytical Ultracentrifugation: Discrete Species and Size-Distributions of Macromolecules and Particles describes the theory and practice of data analysis. Mathematical models for the sedimentation process and the evolution of detected signals are developed in a comprehensive framework, jointly with the description of current and historical strategies for how to extract from noisy experimental data the physical parameters of interest, such as size, mass, and shape, composition, and polydispersity of sedimenting particles. The methods are extensively illustrated, and supported with practical applications, as well as cross-references where to find the methods in the public domain software SEDFIT and SEDPHAT. The systems covered are discrete or polydisperse mixtures of sedimenting molecules or particles in dilute solution, such as proteins and other biomolecules and their stable complexes, man-made polymers, and nanoparticles, observed in different optical systems. These methods form the essential foundation for the analysis of dynamic interacting systems, which are covered in a separate volume.

Analytical ultracentrifugation has become an increasingly important technique for monitoring the size and shape of biological macromolecules. Analytical Ultracentrifugation: Techniques and Methods contains contributions from experts in the field, bringing together the multitude of developments that have taken place in instrumentation and analysis over the past decade into a single volume. This book covers the latest methods in analysis along with an extensive introduction for the novice user. Analysis methods in both sedimentation velocity and sedimentation equilibrium are discussed at length. Protein, protein/DNA, membrane proteins and polymer systems are also explored, along with software developments and non-ideality.

This is the first book of its kind to appear for nearly two decades and gives as comprehensive a coverage as is possible of the present state-of-the-art.

Flow cytometry continually amazes scientists with its ever-expanding utility. Advances in flow cytometry have opened new directions in theoretical science, clinical diagnosis, and medical practice. The new edition of Flow Cytometry: First Principles provides a thorough update of this now classic text, reflecting innovations in the field while outlining the fundamental elements of instrumentation, sample preparation, and data analysis. Flow Cytometry: First Principles, Second Edition explains the basic principles of flow cytometry, surveying its primary scientific and clinical applications and highlighting state-of-the-art techniques at the frontiers of research. This edition contains extensive revisions of all chapters, including new discussions on fluorochrome and laser options for multicolor analysis, an additionalsection on apoptosis in the chapter on DNA, and new chapters onintracellular protein staining and cell sorting, including high-speed sorting and alternative sorting methods, as well as traditional technology. This essential resource: Assumes no prior knowledge of flow cytometry Progresses with an informal, engaging lecture style from simpleto more complex concepts Offers a clear introduction to new vocabulary, principles of instrumentation, and strategies for data analysis Emphasizes the theory relevant to all flow cytometry, with examples from a variety of clinical and scientific fields Flow Cytometry: First Principles, Second Edition provides scientists, clinicians, technologists, and students with the knowledge necessary for beginning the practice of flow cytometry and for understanding related literature.

The third edition of the Encyclopedia of Analytical Science, Ten Volume Set is a definitive collection of articles covering the latest technologies in application areas such as medicine, environmental science, food science and geology. Meticulously organized, clearly written and fully interdisciplinary, the Encyclopedia of Analytical Science, Ten Volume Set provides foundational knowledge across the scope of modern analytical chemistry, linking fundamental topics with the latest methodologies. Articles will cover three broad areas: analytical techniques (e.g., mass spectrometry, liquid chromatography, atomic spectrometry); areas of application (e.g., forensic, environmental and clinical); and analytes (e.g., arsenic, nucleic acids and polycyclic aromatic hydrocarbons), providing a one-stop resource for analytical scientists. Offers readers a one-stop resource with access to information across the entire scope of modern analytical science Presents articles split into three broad areas: analytical techniques, areas of application and and analytes, creating an ideal resource for students, researchers and professionals Provides concise and accessible information that is ideal for non-specialists and readers from undergraduate levels and higher