Enzyme assays are among the most frequently performed procedures in biochemistry and are routinely used to estimate the amount of enzyme present in a cell or tissue, to follow the purification of an enzyme, or to determine the kinetic parameters of a system. The range of techniques used to measure the rate of an enzyme-catalysed reaction is limited only by the nature of the chemical change and the ingenuity of the investigator. This book describes the design and execution of enzyme assays, covering both general principles and specific chapters. Building upon the highly popular first edition, this book combines revised or rewritten chapters with entirely new contributions. Topics include include experimental protocols covering photometric, radiometric, HPLC, and electrochemical assays, along with methods for determining enzyme assays after gel electrophoresis. The theory underlying each method is outlined, together with a description of the instrumentation, sensitivity and sources of error. Also included are chapters on the principles of enzyme assay and kinetic studies; techniques for enzyme extraction; high- throughout screening; statistical analysis of enzyme kinetic data; and the determination of active site concentration. This second edition of Enzyme Assays will be valuable not only to biochemists, but to researchers in all areas of the life sciences.
Biochemistry plays an important role in all areas of the biological and medical sciences. With most of the research or diagnosis involved in these areas being based on biochemically obtained observations, it is essential to have a profile of well standardized protocols. This manual is a basic guide for all students, researchers and experts in biochemistry, designed to help readers in directly starting off their experiments without prior knowledge of the protocol. The book dwells on the concepts used in designing the methodologies, thereby giving ample room for researchers to modify them according to their research requirements.
The two Practical Approach volumes on protein-ligand interaction provide a selection of the most useful and easily applied methods and will be an invaluable guide to the principal techniques used to study the interactions of proteins and ligands. This second volume covers the major spectroscopic methods: FTIR, Raman, and fluorescence spectroscopy; circular dichroism, NMR, mass spectrometry, atomic force microscopy, and the use of paramagnetic probes. There are also chapters on X-ray crystallography and molecular modelling. Hydrodynamic and calorimetric techniques are covered in volume one. Both volumes are written from a practical standpoint to be applicable to both academic and industrial scientists wishing to characterize protein-ligand systems by using a multi-disciplinary approach.
Like the popular first edition, this new edition of Proteolytic Enzymes emphasizes practical aspects of the handling, characterization, inhibition, and use of proteolytic enzymes giving general advice and specific examples. The text and protocols have been thoroughly updated to take account of the advances made in the last 10 years in both the increased understanding of the role of peptidases in many critical cellular processes e.g. apoptosis and new technological developments e.g. in recombinant protein expression, protein sequencing, and structural studies. The topics covered are: nomenclature and classification; purification; assay methods; determination of mechanism; inhibition and prevention of unwanted proteolytic activity; characterizing natural inhibitors; proteolytic enzymes in peptide mapping and primary structure elucidation by mass spectrometry and Edman sequencing; limited proteolysis as a structural probe; synthetic function. This book will be as invaluable as the first edition in providing ideas and protocols for scientists either studying proteases or using proteases as a research tool.
Pregnancy tests, HIV tests, and tests to confirm heart attacks and many other clinical diagnostic tests are all immunoassays. Without immunoassays a huge variety of biologically important molecules could not be easily detected or quantified. Immunoassays will help any life scientist develop his/her own immunoassay.
Immunodiagnostic tests are analytical methods that use antibodies as reagents whose results are used to aid diagnosis and are widely used in many scientific disciplines and in many different ways. Perhaps the most widespread and obvious use is in clinical applications, but immunodiagnostic tests are also used in other fields such as forensic science and environmental and food analysis. The different types of test range from simple manual methods to fully automated systems with sophisticated integrated detection. Immunodiagnostics: A Practical Approach starts off by explaining the principles and development of immunodiagnostic tests, specifically the use of radioisotopes as tracers. Chapter 2 explains the use of solid-phase supports to bind immunoreagents. Enzymes are widely used as labels in immunoassays and their use with colourimetric, fluorimetric, and chemiluminescent detection systems is described. The use of enzymes as labels reflects the move away from radioisotopes and one of the most powerful non- radioisotpoc prodcedures is the time-resolved fluorescence assay. Enzymes can also be used as a simple method of obtaining high performance from immunodiagnostics and this application is covered later in the book. The next set of techniques to be described are light scattering techniques, which can be used in either simple manual assays or in sophisticated automated procedures. The penultimate chapter describes the principles of automation of immunodiagnostic tests. The last topic to be discussed is that of quality assurance.
Practical Hepatic Pathology—a new volume in the new Pattern Recognition series—offers you a practical guide to diagnosing every challenging liver biopsy that you encounter in your daily practice. Dr. Romil Saxena presents diagnoses according to a pattern-based organization that guides you from a histological pattern of injury, through the appropriate work-up, around the pitfalls, and to the best diagnosis. Lavish, full-color images capture key hepatic pathology patterns of injury, pathognomonic features and common variations of all major liver diseases and hepatic neoplasms. No other single source delivers the practical, hands-on information you need to solve even the toughest diagnostic challenges in liver biopsies. Recognize the basic patterns of liver injury through an algorithmic approach and establish diagnosis by a pattern-based visual index present at the beginning of the book. Evaluate and interpret biopsy samples using superb, high-quality, full-color images that illustrate pathognomonic features and common variations. Get comprehensive information on major adult and childhood liver diseases, hepatic neoplasms and pre-neoplastic nodules including clinical features, laboratory tests, imaging findings and differential diagnosis. Understand the pathology and practice of liver transplantation with coverage of the clinical aspects of this procedure.
This book describes the design, construction, optimization and applications of biosensors, including the biological component, its attachment to the transducer, and interfacing of the sensing element to appropriate electronic circuitry. In addition, typical applications of the sensor are described, along with any specimen pretreatment that may be necessary to remove interferences or ensure linearity of the sensor's response. A final chapter on the theoretical analysis of biosensor response characteristics is provided to aid in optimizing any device's performance. Biosensors provides an up-to-date account of techniques which today are being widely used by life scientists in universities and industry.
RNA-protein interactions play a fundamental role in gene expression and protein synthesis. Recent research into the role of RNA in cells has elucidated many more vital interactions with proteins. This book provides an up-to-date and comprehensive guide to a wide range of laboratory procedures to investigate the interactions between RNA and proteins. - ;RNA-protein interactions play a vital role in gene transcription and protein expression. Interactions such as the synthesis of mRNA by RNA polymerases, to the essential modification of RNA by the proteins of the spliceosome complex, and the highly catalytic action of the ribosome in protein synthesis, are established as being fundamental to the function of RNA. Recent research into, for example, the role of RNA as a catalyst, has elucidated many more interactions with proteins that are vital to cell function. RNA - Protein Interactions: A Practical Approach provides a clear and comprehensive guide to the experimental procedures used in studying RNA - protein interactions. The approaches covered range from those initially used to detect a novel RNA-protein interaction, various biochemical and genetic approaches to purifying and cloning RNA binding proteins, through to methods for an in depth analysis of the structural basis of the interaction. The volume includes a number of procedures that have not previously been covered in this type of manual. These include the production of site-specifically modified RNAs by enzymatic and chemical methods and in vivo screening for novel RNA - protein interactions in yeast and E. coli . This is the first volume to gather in one place this wide array of approaches for studying RNA - protein interactions. As is customary for the Practical Approach series, the writing is characterized by a clear explanatory style with many detailed protocols. This informative book will be a valuable aid to laboratory workers in biochemistry and molecular biology - graduate students, postdoctoral and senior scientists - whose research encompasses this field. -