Neuropeptide Y Protocols


Book Description

The observation that neuropeptide Y (NPY) is the most abundant peptide present in the mammalian nervous system and the finding that it elicits the most powerful orexigenic signal have led to active investigations of the properties of the NPY family of hormones, including peptide YY (PYY) and pancreatic polypeptide (PP). Nearly two decades of research have led to the identification of several NPY receptor subtypes and the development of useful receptor selective ligands. Moreover, these investigations have imp- cated NPY in the pathophysiology of a number of diseases, including feeding disorders, seizures, memory loss, anxiety, depression, and heart failure. Vigorous efforts are therefore continuing, not only to understand the bioche- cal aspects of NPY actions, but also toward developing NPY-based treatments for a variety of disorders. To facilitate these efforts, it was decided to produce the first handbook on NPY research techniques as part of the Methods in Molecular Biology Series. In compiling Neuropeptide Y Protocols, I have gathered contributions on techniques considered critical for the advancement of the NPY field from experts in various disciplines. Each chapter starts with a brief introduction, with Materials and Methods sections following. The latter sections are presented in an easy to follow step-by-step format. The last section of the chapter, Notes, highlights pitfalls and the maneuvers employed to overcome them. This information, not usually disseminated in standard research pub- cations, may prove extremely useful for investigators employing these te- niques in NYP research.




Capillary Electrophoresis of Nucleic Acids


Book Description

The development of PCR, which enables extremely small amounts of DNA to be amplified, led to the rapid development of a multiplicity of a- lytical procedures that permit use of this new resource for the analysis of genetic variation and for the detection of disease-causing mutations. The advent of capillary electrophoresis (CE), with its power to separate and a- lyze very small amounts of DNA, has also stimulated researchers to develop analytical procedures for the CE format. The advantages of CE in terms of speed and reproducibility of analyses are manifold. Furthermore, the high s- sitivity of detection, and the ability to increase sample throughput with par- lel analysis, has led to the creation of a full range of analysis of DNA molecules, from modified DNA adducts and single-strand oligonucleotides through PCR-amplified DNA fragments and whole chromosomes. Capillary Elect- phoresis of Nucleic Acids focuses on analytical protocols that can be used for detection and analysis of mutations and modification, from precise DNA loci through entire genomes of organisms. Important practical considerations for CE, such as the choice of separation media, electrophoresis conditions, and the influence of buffer additives and dyes on DNA mobility, are discussed in several key chapters and within particular applications.




Protein Structure, Stability, and Folding


Book Description

In Protein Structure, Stability, and Folding, Kenneth P. Murphy and a panel of internationally recognized investigators describe some of the newest experimental and theoretical methods for investigating these critical events and processes. Among the techniques discussed are the many methods for calculating many of protein stability and dynamics from knowledge of the structure, and for performing molecular dynamics simulations of protein unfolding. New experimental approaches presented include the use of co-solvents, novel applications of hydrogen exchange techniques, temperature-jump methods for looking at folding events, and new strategies for mutagenesis experiments. Unique in its powerful combination of theory and practice, Protein Structure, Stability, and Folding offers protein and biophysical chemists the means to gain a more comprehensive understanding of some of this complex area by detailing many of the major techniques in use today.




The ELISA Guidebook


Book Description

John R. Crowther provides today's premier practical guide to the understanding and application of ELISA. Updating and greatly expanding his widely appreciated earlier publication, ELISA Theory and Practice (1995), this important work introduces chapters on such major new topics as checkerboard titrations, quality control of testing, kit production and control, novel monoclonal antibodies, validation of assays, statistical requirements for data examination, and epidemiological considerations. With its numerous worked examples, detailed instructions, and extensive illustrations, The ELISA Guidebook offers a powerful synthesis of all the basic concepts and practical experimental details investigators need to understand, develop, and apply the new ELISA methodology successfully in day-to-day basic and clinical research.




In Situ Hybridization Protocols for the Brain


Book Description

This volume of the International Review of Neurobiology was written to assist researchers without any previous experience with in situ hybridization, allowing them to follow the protocols and expect good results. It contains all the information required for newcomers to achieve successful in situ hybridization results, and methods for improving the technique of those already utilizing it. Published since 1959, International Review of Neurobiology is a well-known series appealing to neuroscientists, clinicians, psychologists, physiologists, and pharmacologists. Led by an internationally renowned editorial board, this important serial publishes both eclectic volumes made up of timely reviews and thematic volumes that focus on recent progress in a specific area of neurobiology research. A well-known series appealing to neuroscientists, clinicians, psychologists, physiologists, and pharmacologists Led by an internationally renowned editorial board, this important serial publishes both eclectic volumes made up of timely reviews and thematic volumes that focus on recent progress in a specific area of neurobiology research




Biostatistical Methods


Book Description

Leading biostatisticians and biomedical researchers describe many of the key techniques used to solve commonly occurring data analytic problems in molecular biology, and demonstrate how these methods can be used in the development of new markers for exposure to a risk factor or for disease outcomes. Major areas of application include microarray analysis, proteomic studies, image quantitation, genetic susceptibility and association, evaluation of new biomarkers, and power analysis and sample size.




Posttranslational Modification of Proteins


Book Description

Christoph Kannicht and a panel of highly experienced researchers describe readily reproducible methods for detecting and analyzing the posttranslational modifications of protein, particularly with regard to protein function, proteome research, and the characterization of pharmaceutical proteins.




Handbook of Endocrine Research Techniques


Book Description

Written by experts in the field, Handbook of Endocrine Research Techniques is currently the only single source of up-to-date methods and strategies particularly useful in endocrinological research. As a resource for both the novice and experienced investigator, the book includes chapters which provide an introduction to the area, general concepts, detailed protocols, and extensive references.







Adipose Tissue Protocols


Book Description

Adipose tissue is recognized to be exquisitely sensitive to hormone action, and is also now recognized as a secretory and endocrine organ required for reproduction and good health. Adipocytes are “smart” cells able within the tissue to communicate with surrounding cells, but also with various organs, particularly via leptin acting on the central nervous system. Brown adipose tissue (BAT) and white adipose tissue (WAT) are known to be distinct tissues, whereas the heterogeneity of WAT depots is well est- lished. Unfortunately, excess WAT leads to obesity, which is the most common health problem in industrialized countries. Therefore, from both a scientific and a technical point of view, the time has come to create a survey of adipose tissues and their neglected adipocytes. In Adipose Tissue Protocols, I have attempted to gather together chapters from all areas of adipose tissue research—from in vivo to in vitro studies—and to provide methods covering a wide variety of techniques, including the choice of adipose tissue depot and of morphological techniques for the study of BAT and WAT; the isolation, subcellular fractionation, and transfection of adipocytes where the low density of these cells must be taken into account; assays of nutrient and ion fluxes and the metabolic aspects of nutrient uptake; assays of lipid-related enzymes; biopsies and quantification of lipid-related mRNAs; cultures of adipose precursor cells from WAT and BAT of various species, including human tissue; measurements of adipose secretory products; and assessment of WAT metabolism in vivo.