This volume describes protocols for basic state-of-the-art approaches in the field of peptidomics. Most of these approaches are independent of the instruments used for analysis and can easily be adapted for equipment that is available in a typical proteomics facility. Chapters detail many of the basic techniques used to detect and identify peptides, methods for the relative quantitation of peptides between samples using isotopic labels or label-free approaches, and biological species as well as sample types. Written in the highly successful format of the Methods in Molecular Biology series, each chapter includes an introduction to the topic, a list of the necessary materials and reagents, reproducible step-by-step laboratory protocols, and tips on troubleshooting common problems and avoiding pitfalls. Authoritative and practical, Peptidomics: Methods and Strategies provides useful guidance for studies in the rapidly growing field of peptidomics.

Transport and Diffusion across Cell Membranes is a comprehensive treatment of the transport and diffusion of molecules and ions across cell membranes. This book shows that the same kinetic equations (with appropriate modification) can describe all the specialized membrane transport systems: the pores, the carriers, and the two classes of pumps. The kinetic formalism is developed step by step and the features that make a system effective in carrying out its biological role are highlighted. This book is organized into six chapters and begins with an introduction to the structure and dynamics of cell membranes, followed by a discussion on how the membrane acts as a barrier to the transmembrane diffusion of molecules and ions. The following chapters focus on the role of the membrane's protein components in facilitating transmembrane diffusion of specific molecules and ions, measurements of diffusion through pores and the kinetics of diffusion, and the structure of such pores and their biological regulation. This book methodically introduces the reader to the carriers of cell membranes, the kinetics of facilitated diffusion, and cotransport systems. The primary active transport systems are considered, emphasizing the pumping of an ion (sodium, potassium, calcium, or proton) against its electrochemical gradient during the coupled progress of a chemical reaction while a conformational change of the pump enzyme takes place. This book is of interest to advanced undergraduate students, as well as to graduate students and researchers in biochemistry, physiology, pharmacology, and biophysics.

This detailed volume provides in-depth protocols for protein labeling techniques and applications, with an additional focus on general background information on the design and generation of the organic molecules used for the labeling step. Chapters provide protocols for labeling techniques and applications, with an additional focus on general background information on the design and generation of the organic molecules used for the labeling step. 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 practical, Site-Specific Protein Labeling: Methods and Protocols provides a comprehensive overview on the most relevant and established labeling methodologies, and helps researchers to choose the most appropriate labeling method for their biological question.

This book is designed to be a practical progression of experimental techniques an investigator may follow when embarking on a biochemical project. The protocols may be performed in the order laid out or may be used inde pendently. The aim of the book is to assist a wide range of researchers. from the novice to the frustrated veteran, in the choice and design of experiments that are to be performed to provide answers to specific questions. The manual describes standard techniques that have been shown to work, as well as some newer ones that are beginning to prove important. By following the promi nently numbered steps. you can work your way through any protocol. whether it's a new technique or a task you've done before for which you need a quick review or updated methodology. This manual will assist the experimentalist in designing properly controlled experiments. There will be no advice for dealing with specific pieces of equip ment other than encouragement to read the manual, if you can find it. Through out all manipulations try to be objective. Be on the lookout for unexpected findings. You will learn the most from unexpected results. and they are often the beginning of the next project. It is never possible to record too much in your lab notebook. Do not get discouraged. Remember, things will not always run smoothly.

Describes experimental methods for investigating the function of pumps, channels and transporters Covers new emerging analytical methods used to study ion transport membrane proteins such as single-molecule spectroscopy Details a wide range of electrophysiological techniques and spectroscopic methods used to analyze the function of ion channels, ion pumps and transporters Covers state-of-the art analytical methods to study ion pumps, channels, and transporters, and where analytical chemistry can make further contributions