Download or read book Kinesin Protocols written by Isabelle Vernos and published by Humana. This book was released on 2010-11-10 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: By the end of the 1980s only two microtubule-dependent motors, the plus end-directed kinesin and the minus end-directed cytoplasmic dynein, had been identified. At the time, these two motors seemed almost sufficient to explain directional motility events on polar microtubule tracks in the cell. No- theless, shortly after, the tip of the iceberg began to emerge with the identi- cation of proteins containing in their sequences a domain found in kinesin. This domain, called the “motor domain,” conferred on these proteins the essential property of moving on microtubules, using the energy derived from ATP hydro- sis. Since then, the identification of new proteins belonging to the kinesin superfamily of microtubule-dependent motors has gone at such a pace that nowadays more than 200 entries with motor domain sequences are deposited in the database. Kinesin family members are found in all eukaryotic org- isms tested. They present a wide range of domain organizations with a motor domain located at different positions in the molecule. Their motility prop- ties are also variable in directionality, velocity, and such other characteristics as bundling activity and processivity. Finally, and most important, they p- ticipate in a multitude of cellular functions. Our understanding of many cel- lar events, such as mitotic spindle assembly and neuronal transport, to cite only two, has progressed substantially in the last few years thanks to the id- tification of these motors.

Download or read book Microtubule Protocols written by Jun Zhou and published by Springer Science & Business Media. This book was released on 2007-08-23 with total page 307 pages. Available in PDF, EPUB and Kindle. Book excerpt: Microtubules are essential components of the cytoskeleton, and play critical roles in a variety of cell processes, including cell shaping, intracellular tracking, cell division, and cell migration. Microtubule Protocols presents a comprehensive collection of essential and up-to-date methods for studying both the biology of microtubules and the mechanisms of action of microtubule-interacting drugs. The straightforward presentation of readily reproducible protocols is a hallmark of the Methods in Molecular MedicineTM series, and is evident in this volume. Methods presented range from the purification and characterization of microtubule proteins, analysis of post-translational modifications of tubulin, and determination of microtubule structure, to the visualization of microtubule and spindle behavior, measurement of microtubule dynamics, and examination of microtubule-mediated cellular processes. Both basic scientists and clinical researchers will benefit from this collection of state-of-the-art protocols for microtubule research.

Download or read book Connexin Methods and Protocols written by Roberto Bruzzone and published by Springer Science & Business Media. This book was released on 2008-02-05 with total page 495 pages. Available in PDF, EPUB and Kindle. Book excerpt: Direct cell–cell communication is a common property of multicellular organisms that is achieved through membrane channels which are organized in gap junctions. The protein subunits of these intercellular channels, the connexins, form a multigene family that has been investigated in great detail in recent years. It has now become clear that, in different tissues, connexins speak several languages that control specific cellular functions. This progress has been made possible by the availability of new molecular tools and the improvement of basic techniques for the study of membrane channels, as well as by the use of genetic approaches to study protein function in vivo. More important, connexins have gained visibility because mutations in some connexin genes have been found to be linked to human genetic disorders. Connexin Methods and Protocols presents in detail a collection of te- niques currently used to study the cellular and molecular biology of connexins and their physiological properties. The field of gap junctions and connexin research has always been characterized by a multidisciplinary approach c- bining morphology, biochemistry, biophysics, and cellular and molecular biology. This book provides a series of cutting-edge protocols and includes a large spectrum of practical methods that are available to investigate the fu- tion of connexin channels. Connexin Methods and Protocols is divided into three main parts.

Download or read book Cytoskeleton Methods and Protocols written by Ray H. Gavin and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 286 pages. Available in PDF, EPUB and Kindle. Book excerpt: Over the past two decades experimental studies have solidified the int- pretation of the cytoskeleton as a highly dynamic network of microtubules, actin microfilaments, intermediate filaments, and myosin filaments. Rather than a network of disparate fibers, these polymers are often interconnected and display synergy, which is the combined action of two or more cytoskeletal polymers to achieve a specific cellular structure or function. Cross-commu- cation among cytoskeletal polymers is thought to be achieved through cytoskeletal polymer accessory proteins and molecular motors that bind two or more cytoskeletal polymers. Development of the modern concept of the cytoskeleton is a direct o- growth of advances in experimental tools and reagents that are available to cell and molecular biologists. Technological advances and refinements in cell imaging have made it possible to selectively image a single cytoskeletal po- mer and monitor its dynamics through the use of fluorescence probes in vitro and in vivo. Two decades ago, cytoskeletal research was limited to a few perturbation reagents that included colchicine and cytochalasin. Today, the perturbation arsenal has expanded to a highly selective group of reagents that includes Taxol, nocodazole, benomyl, latrunculin, jasplakinolide, and such endogenous proteins as gelsolin. These reagents enable the investigator to selectively perturb or destroy a cytoskeletal polymer while leaving other cytoskeletal polymers intact. Site-specific monoclonal antibodies that target a specific cytoskeletal polymer have proven to be highly selective affinity tools for cytoskeletal research.

Download or read book Nuclease Methods and Protocols written by Catherine H. Schein and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 521 pages. Available in PDF, EPUB and Kindle. Book excerpt: Nucleases, enzymes that restructure or degrade nucleic acid polymers, are vital to the control of every area of metabolism. They range from “housekeeping” enzymes with broad substrate ranges to extremely specific tools (1). Many types of nucleases are used in lab protocols, and their commercial and clinical uses are expanding. The purpose of Nuclease Methods and Protocols is to introduce the reader to some we- characterized protein nucleases, and the methods used to determine their activity, structure, interaction with other molecules, and physiological role. Each chapter begins with a mini-review on a specific nuclease or a nuclease-related theme. Although many chapters cover several topics, they were arbitrarily divided into five parts: Part I, “Characterizing Nuclease Activity,” includes protocols and assays to determine general (processive, distributive) or specific mechanisms. Methods to assay nuclease products, identify cloned nucleases, and determine their physiological role are also included here. Part II, “Inhibitors and Activators of Nucleases,” summarizes assays for measuring the effects of other proteins and small molecules. Many of these inhibitors have clinical relevance. Part III, “Relating Nuclease Structure and Function,” provides an overview of methods to determine or model the 3-D structure of nucleases and their complexes with substrates and inhibitors. A 3-D structure can greatly aid the rational design of nucleases and inhibitors for specific purposes. Part IV, “Nucleases in the Clinic,” summarizes assays and protocols suitable for use with t- sues and for nuclease based therapeutics.

Download or read book Genomics Protocols written by Michael P. Starkey and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 537 pages. Available in PDF, EPUB and Kindle. Book excerpt: We must unashamedly admit that a large part of the motivation for editing Genomics Protocols was selfish. The possibility of assembling in a single volume a unique and comprehensive collection of complete protocols, relevant to our work and the work of our colleagues, was too good an opportunity to miss. We are pleased to report, however, that the outcome is something of use not only to those who are experienced practitioners in the genomics field, but is also valuable to the larger community of researchers who have recognized the potential of genomics research and may themselves be beginning to explore the technologies involved. Some of the techniques described in Genomics Protocols are clearly not restricted to the genomics field; indeed, a prerequisite for many procedures in this discipline is that they require an extremely high throughput, beyond the scope of the average investigator. However, what we have endeavored here to achieve is both to compile a collection of procedures concerned with geno- scale investigations and to incorporate the key components of “bottom-up” and “top-down” approaches to gene finding. The technologies described extend from those traditionally recognized as coming under the genomics umbrella, touch on proteomics (the study of the expressed protein complement of the genome), through to early therapeutic approaches utilizing the potential of genome programs via gene therapy (Chapters 27–30).

Download or read book Immunotoxin Methods and Protocols written by Walter A. Hall and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 305 pages. Available in PDF, EPUB and Kindle. Book excerpt: Immunotoxins represent a new class of human therapeutics that have widespread applications and a potential that has not yet been fully recognized since they were first conceived of by Paul Ehrlich in 1906. The majority of advances in the development and implementation of immunotoxins has occurred over the last 20 years. The reasons for this use of immunotoxins in basic science and clinical research are the powerful concurrent advances in genetic engineering and receptor physiology. Recombinant technology has allowed investigators to produce sufficient quantities of a homogeneous c- pound that allows clinical trials to be performed. The identification of specific receptors on malignant cell types has enabled scientists to generate immunotoxins that have had positive results in clinical trials. As more cellular targets are identified in coming years, additional trials will be conducted in different disease states affecting still larger patient populations. Modulation of the immune system to decrease the humoral response to immunotoxins may improve their overall efficacy. As increasingly more effective compounds are generated, it will be necessary to decrease the local and systemic toxicity - sociated with these agents, and methods for doing so are presently being - veloped. The work presented in Immunotoxin Methods and Protocols focuses on three specific areas of immunotoxin investigation that are being conducted by experts throughout the world. The first section describes the construction and development of a variety of immunotoxins.

Download or read book Neuropeptide Y Protocols written by Ambikaipakan Balasubramaniam and published by Springer Science & Business Media. This book was released on 2008-02-05 with total page 244 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.

Download or read book Neurotrophin Protocols written by Robert A. Rush and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 272 pages. Available in PDF, EPUB and Kindle. Book excerpt: The past decade has seen an extraordinary growth in research interest in neurotrophic factors, and the study of the neurotrophin family has led this activity. Nevertheless, this area of research has often struggled as a result of techniques that were either inadequate or just emerging from other research fields and disciplines. Neurotrophin Protocols has brought together many leaders in the neurotrophin field who detail their special expertise in a wide variety of techniques. Though most procedures are valid across many diff- ent fields of research, some of those described here have been developed to address particular issues within the neurotrophic factor field. The protocols cover a broad range of biochemical, histological, and biological techniques that are often required by the modern laboratory. However, all have been written with sufficient detail to allow any laboratory to achieve proficiency without need of reference to other texts. Neurotrophin Protocols is divided into four sections dealing with p- tein, RNA, recombinant, and in vivo techniques. Protein techniques have in general been less successfully employed than those dealing with RNA or DNA. However, procedures that achieve localization and quantification of the neurotrophins are now being used more extensively. Their inclusion here should assist further studies at the protein level. Transgenic cell lines and animals are commonplace in the scientific research literature, but their inc- sion in several chapters in this book provide some novel uses that are not readily available elsewhere.

Download or read book Mycotoxin Protocols written by Mary W. Trucksess and published by Springer Science & Business Media. This book was released on 2008-02-05 with total page 245 pages. Available in PDF, EPUB and Kindle. Book excerpt: Mycotoxins produced by molds are common contaminants of many important crops, including wheat, corn, rice, and peanuts. Some mycotoxins are found in fruits and vegetables. These contaminants have a broad range of toxic effects, including carcinogenicity, neurotoxicity, and reproductive and developmental toxicity. The occurrence of mycotoxins in foods is an unavoidable worldwide problem. About 80 countries have imposed regulatory limits to minimize human and animal exposure to mycotoxins. Regulatory limits, including international standards, have tremendous economic impact and must be developed using science-based risk assessments. The purpose of Mycotoxin Protocols is to provide the scientific and technological basis for analytical methods for use in obtaining the exposure data needed for risk assessments. Mycotoxin Protocols is divided into four sections, which are interc- nected. The first section: Chapters 1–5 describe the general techniques for mycotoxin analysis with emphasis on the importance of method validation based on statistical parameters; sampling procedures for collecting a sample as representative as possible of a bulk lot; the isolation of mycotoxins for use as analytical standards or for toxicological studies; the evaluation of purity and preparation of standards; and the detection and identification of impu- ties in isolated mycotoxins. Sections 2–4: Chapters 6–19 describe the most current chromatographic and immunochemical methods for studies on the major mycotoxins.

Download or read book The Kinesin Superfamily Handbook written by Claire T. Friel and published by CRC Press. This book was released on 2020-05-13 with total page 132 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book brings together current information on the families that make up the kinesin superfamily of molecular motors in one comprehensive text; an ideal reference for researchers looking to make comparisons between different families, for specific information on an individual family, or simply for an overview of the kinesin superfamily. Information is clearly structured and grouped according to individual families and organised in a standardised way, allowing the reader to easily search and retrieve information on this large superfamily of molecular motors and understand how its individual members carry out a diverse variety of cellular functions. Features : The first book dedicated to the entire kinesin superfamily Enables a fuller understanding of this family of proteins, which is becoming an increasing focus of research due to its involvement in diseases such as cancer, neuropathies and ciliopathies Written in a manner accessible to a range of researchers in the life and medical sciences, including biophysicists, biochemists and medical researchers. Chapters 4 and 11 of this book are freely available as downloadable Open Access PDFs at http://www.taylorfrancis.com under a Creative Commons Attribution-Non Commercial-No Derivatives (CC-BY-NC-ND) 4.0 license.

Download or read book Proteoglycan Protocols written by Renato V. Iozzo and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 547 pages. Available in PDF, EPUB and Kindle. Book excerpt: Proteoglycans are some of the most elaborate macromolecules of mammalian and lower organisms. The covalent attachment of at least five types of glycosami- glycan side chains to more than forty individual protein cores makes these molecules quite complex and endows them with a multitude of biological functions. Proteoglycan Protocols offers a comprehensive and up-to-date collection of prepa- tive and analytical methods for the in-depth analysis of proteoglycans. Featuring st- by-step detailed protocols, this book will enable both novice and experienced researchers to isolate intact proteoglycans from tissues and cultured cells, to establish the composition of their carbohydrate moieties, to generate strategies for prokaryotic and eukaryotic expression, to utilize methods for the suppression of specific proteoglycan gene expression and for the detection of mutant cells and degradation products, and to study specific interactions between proteoglycans and extracellular matrix proteins as well as growth factors and their receptors. The readers will find concise, yet comprehensive techniques carefully drafted by leading experts in the field. Each chapter commences with a general Introduction, followed by a detailed Materials section, and an easy-to-follow Methods section. An asset of each chapter is the extensive notation that includes troubleshooting tips and practical considerations that are often lacking in formal methodology papers. The reader will find this section most valuable because it is clearly provided by experienced scientists who have first-hand knowledge of the techniques they outline. In addition, most of the chapters are well illustrated with examples of typical data generated with each method.

Download or read book Adipose Tissue Protocols written by Gérard Ailhaud and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 332 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.

Download or read book Gene Knockout Protocols written by Martin J. Tymms and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 434 pages. Available in PDF, EPUB and Kindle. Book excerpt: As the major task of sequencing the human genome is near completion and full complement of human genes are catalogued, attention will be focused on the ultimate goal: to understand the normal biological functions of these genes, and how alterations lead to disease states. In this task there is a severe limitation in working with human material, but the mouse has been adopted as the favored animal model because of the available genetic resources and the highly conserved gene conservation linkage organization. In just of ten years since the first gene-targeting experiments were p- formed in embryonic stem (ES) cells and mutations transmitted through the mouse germline, more than a thousand mouse strains have been created. These achievements have been made possible by pioneering work that showed that ES cells derived from preimplantation mouse embryos could be cultured for prolonged periods without differentiation in culture, and that homologous rec- bination between targeting constructs and endogenous DNA occurred at a f- quency sufficient for recombinants to be isolated. In the next few years the mouse genome will be systematically altered, and the techniques for achi- ing manipulations are constantly being streamlined and improved.

Download or read book Amino Acid Analysis Protocols written by Catherine Cooper and published by Springer Science & Business Media. This book was released on 2008-02-05 with total page 268 pages. Available in PDF, EPUB and Kindle. Book excerpt: A collection of classic and cutting-edge techniques of high utility in answering specific biological questions about amino acids. Common methods include those based on HPLC or gas chromatography separation and analysis after precolumn derivatization. New techniques based on capillary electrophoresis separation, high-performance anion exchange chromatography, and mass spectrometry are also presented. Each method is described in step-by-step detail to ensure successful experimental results and emphasizes sample preparation, particularly the collection and storage of bodily fluids. Up-to-date and highly practical, Amino Acid Analysis Protocols offers analytical and clinical chemists, as well as a broad range of biological and biomedical investigators, a rich compendium of laboratory tools for the productive analysis of both common and uncommon amino acids.

Download or read book GTPase Protocols written by Ed Manser and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 259 pages. Available in PDF, EPUB and Kindle. Book excerpt: In the last 10 years researchers have firmly established key roles for R- related GTPases in almost every aspect of cell biology. In the 1980s the pro- oncogene Ras itself was the focus of interest, though in the 1990s this shifted to the increasing variety of Ras-related proteins. In this new decade much yet needs to be done to establish the role for all the small GTPases now uncovered by the human genome project. In particular, these GTPases need to be und- stood in the appropriate biochemical and cellular contexts. In the process of trying to uncover the role of these versatile proteins, a variety of novel te- niques and methodologies has been developed. These now enable investi- tors to move easily within a diversity of fields ranging from structural studies to real-time in vivo analysis of a GTPase. In recognition of the need for access to key background methodologies, GTPase Protocols: The Ras Superfamily is devoted to techniques that are pr- ently widely used and that will continue to be the standard for researchers worldwide. Each chapter is aimed at supplying detailed methodologies to allow reproduction in any laboratory, while also providing the general pr- ciples on which the methods are based. Some of the techniques grouped in the first section apply broadly to small GTPases, whereas others in Part II are more applicable within each GTPase subfamily.

Download or read book Matrix Metalloproteinase Protocols written by Ian M. Clark and published by Springer Science & Business Media. This book was released on 2008-02-05 with total page 547 pages. Available in PDF, EPUB and Kindle. Book excerpt: Research in the matrix metalloproteinase field began with the demonstration by Gross and Lapière, in 1962, that resorbing tadpole tail expressed an enzyme that could degrade collagen gels. These humble beginnings have led us to the elucidation of around twenty distinct vertebrate MMPs, along with a variety of homologs from such diverse organisms as sea urchin, plants, nematode worm, and bacteria. This, coupled with four known specific inhibitors of MMPs, the TIMPs, gives a complex picture. Part I of Matrix Metalloproteinase Protocols provides the reader with a selective overview of the MMP arena, and a chance to come to grips with where the field has been, where it is, and where it is going. I hope that this complements all of the methodology that comes later. Part II presents the reader with a diverse set of methods for the expression and purification of MMPs and TIMPs, bringing together the long and often hard-earned experience of a number of researchers. Part III allows the reader to detect MMPs and TIMPs at both the protein and mRNA level, whereas Part IV gives the ability to assay MMP and TIMP activities in a wide variety of circumstances.