Neurotrophin Protocols

Author : Robert A. Rush
Publisher : Springer Science & Business Media
Page : 272 pages
File Size : 55,9 Mb
Release : 2008-02-03
Category : Medical
ISBN : 9781592590605

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Neurotrophin Protocols by Robert A. Rush Pdf

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.

Author : Stephen D Skaper
Publisher : Humana Press
Page : 421 pages
File Size : 45,8 Mb
Release : 2012-02-25
Category : Medical
ISBN : 1617795372

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Neurotrophic Factors by Stephen D Skaper Pdf

Nervous system development evolves from the well-orchestrated processes of neural induction, cell proliferation, differentiation, cell migration, survival, and synapse formation. Among these environmental cues, neurotrophic factors are secreted proteins that promote neurite outgrowth, neuronal cell differentiation and survival both in vivo and in vitro. Nerve growth factor (NGF) is the founding and best characterized member of the neurotrophin family of neurotrophic polypeptides. Neurotrophic Factors: Methods and Protocols presents a selection of protocols and procedures which make use of cellular, tissue, and whole animal models which can be applied to the investigation of neurotrophic factors and other agents impacting on these systems. Chapters cover a wide-range of topics such as dealing with the culture of neurons and glia from the central and peripheral nervous systems, neuron-glia co-culture models, and cell-based assays for the evaluation of neuroprotective molecules, as well as assays which can be applied to the study of agents with neuroregenerative potential. Protocols describing viral- and nanoparticle-based delivery methods to neural cells are also presented, following by chapters dealing with organotypic slice culture protocols. Lastly, several chapters are dedicated to in vivo lesion models of relevance to nervous system pathology, which can be applied to the investigation of neurotrophic factors and peptides. Written in the successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Neurotrophic Factors: Methods and Protocols seeks to serve both professionals and novices alike with its well-honed methodologies in an effort to further our knowledge of what has been described as the last frontier of science.

Author : John M. Walker
Publisher : Unknown
Page : 128 pages
File Size : 54,7 Mb
Release : 1984
Category : Molecular biology -- v.236. Plant functional genomics -- v.237. G protein signaling -- v.238. Biopolymer methods in tissue engineering -- v.239. Cell migration in inflammation and immunity -- v.240. Mammalian artificial chromosomes -- v.241. Cell cycle checkpoint control protocols -- v.242. Atomic force microscopy -- v.243. Chiral separations: Methods and protocols -- v.244. Protein purification protocols -- v.245-6. Gene delivery to mammalian cells -- v.247. Drosophila cytogenetics protocols -- v.248. Antibody engineering -- v.249. Cytokine protocols -- v.251. HPLC of peptides and proteins: Methods and protocols -- v.265. RNA interference, editing, and modification -- v.274. Photosynthesis research protocols -- v.318. Plant cell culture protocols -- v.323. Arabidopsis protocols
ISBN : LCCN:84015696

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Methods in Molecular Biology: Neurotrophin protocols by John M. Walker Pdf

Author : Robert A. Rush
Publisher : Humana Press
Page : 274 pages
File Size : 46,5 Mb
Release : 2000-11-07
Category : Medical
ISBN : 0896036995

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Neurotrophin Protocols by Robert A. Rush Pdf

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.

Author : Renato V. Iozzo
Publisher : Springer Science & Business Media
Page : 547 pages
File Size : 49,8 Mb
Release : 2008-02-02
Category : Science
ISBN : 9781592592098

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Proteoglycan Protocols by Renato V. Iozzo Pdf

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.

Author : Michael P. Starkey,Ramnath Elaswarapu
Publisher : Springer Science & Business Media
Page : 537 pages
File Size : 55,6 Mb
Release : 2008-02-03
Category : Medical
ISBN : 9781592592357

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Genomics Protocols by Michael P. Starkey,Ramnath Elaswarapu Pdf

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).

Author : Gérard Ailhaud
Publisher : Springer Science & Business Media
Page : 332 pages
File Size : 46,7 Mb
Release : 2008-02-03
Category : Science
ISBN : 9781592592319

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Adipose Tissue Protocols by Gérard Ailhaud Pdf

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.

Author : Mary W. Trucksess,Albert E. Pohland
Publisher : Springer Science & Business Media
Page : 245 pages
File Size : 54,6 Mb
Release : 2008-02-05
Category : Medical
ISBN : 9781592590643

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Mycotoxin Protocols by Mary W. Trucksess,Albert E. Pohland Pdf

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.

Author : Martin J. Tymms,Ismail Kola
Publisher : Springer Science & Business Media
Page : 434 pages
File Size : 46,5 Mb
Release : 2008-02-03
Category : Medical
ISBN : 9781592592203

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Gene Knockout Protocols by Martin J. Tymms,Ismail Kola Pdf

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.

Author : Ian M. Clark
Publisher : Springer Science & Business Media
Page : 547 pages
File Size : 52,7 Mb
Release : 2008-02-05
Category : Science
ISBN : 9781592590469

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Matrix Metalloproteinase Protocols by Ian M. Clark Pdf

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.

Author : Neil Osheroff,Mary-Ann Bjornsti
Publisher : Springer Science & Business Media
Page : 331 pages
File Size : 42,8 Mb
Release : 2008-02-05
Category : Medical
ISBN : 9781592590575

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DNA Topoisomerase Protocols by Neil Osheroff,Mary-Ann Bjornsti Pdf

Beginning with the Escherichia coli ? protein, or bacterial DNA topoisomerase I, an ever-increasing number of enzymes have been identified that catalyze changes in the linkage of DNA strands. DNA topoisomerases are ubiquitous in nature and have been shown to play critical roles in most p- cesses involving DNA, including DNA replication, transcription, and rec- bination. These enzymes further constitute the cellular targets of a number of clinically important antibacterial and anticancer agents. Thus, further studies of DNA topology and DNA topoisomerases are critical to advance our und- standing of the basic biological processes required for cell cycle progression, cell division, genomic stability, and development. In addition, these studies will continue to provide critical insights into the cytotoxic action of drugs that target DNA topoisomerases. Such mechanistic studies have already played an important role in the development and clinical application of antimicrobial and chemotherapeutic agents. The two volumes of DNA Topoisomerase Protocols are designed to help new and established researchers investigate all aspects of DNA topology and the function of these enzymes. The chapters are written by prominent investigators in the field and provide detailed background information and st- by-step experimental protocols. The topics covered in Part I: DNA Topology and Enzymes, range from detailed methods to analyze various aspects of DNA structure, from linking number, knotting/unknotting, site-specific recombi- tion, and decatenation to the overexpression and purification of bacterial and eukaryotic DNA topoisomerases from a variety of cell systems and tissues.

Author : Clare Wise
Publisher : Springer Science & Business Media
Page : 407 pages
File Size : 53,6 Mb
Release : 2008-02-05
Category : Medical
ISBN : 9781592591855

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Epithelial Cell Culture Protocols by Clare Wise Pdf

There have been significant advances in research involving the isolation and culture of epithelial cells in the past decade, and many new techniques have been developed. Monolayer cultures can be used to evaluate the nature and behavior of cells, while the use of epithelial cells in model systems has allowed a deeper understanding of cellular and molecular mechanisms and interactions. The aim of this book is to provide a comprehensive, step-by-step guide to many techniques for epithelial cell culture, combining in one volume the more commonly used protocols along with many that are more speci- ized. Epithelial Cell Culture Protocols should help those who are new to this field and want to learn the basic culture techniques, as well as those needing to use more wide ranging and specific protocols. It should be a useful resource on its own, and also complement the other volumes that have been written about cell culture in the Methods in Molecular Biology series. Epithelial Cell Culture Protocols covers a wide variety of protocols, mostly aimed at the researcher, but also a few aimed at clinicians. The est- lishment and maintenance of primary cultures derived from many different tissues and different species is covered. Particular emphasis has been placed on protocols needed to further analyze and assess epithelial cells, for example, by looking at apoptosis and integrins and by measuring membrane capa- tance and confluence. Using different co-culture techniques, it is possible also to develop models to investigate many different systems in vitro.

Author : Ray H. Gavin
Publisher : Springer Science & Business Media
Page : 286 pages
File Size : 44,5 Mb
Release : 2008-02-03
Category : Science
ISBN : 9781592590513

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Cytoskeleton Methods and Protocols by Ray H. Gavin Pdf

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.

Author : Walter A. Hall
Publisher : Springer Science & Business Media
Page : 305 pages
File Size : 46,7 Mb
Release : 2008-02-02
Category : Medical
ISBN : 9781592591145

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Immunotoxin Methods and Protocols by Walter A. Hall Pdf

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.

Author : Roberto Bruzzone,Christian Giaume
Publisher : Springer Science & Business Media
Page : 495 pages
File Size : 52,8 Mb
Release : 2008-02-05
Category : Science
ISBN : 9781592590438

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Connexin Methods and Protocols by Roberto Bruzzone,Christian Giaume Pdf

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.