Connexin Methods And Protocols
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Connexin Methods And Protocols
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Author : Roberto Bruzzone
language : en
Publisher: Springer Science & Business Media
Release Date : 2008-02-05
Connexin Methods And Protocols written by Roberto Bruzzone and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2008-02-05 with Science categories.
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.
Connexin Hemichannels
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Author : Fabio Mammano
language : en
Publisher: Springer Nature
Release Date : 2024-04-05
Connexin Hemichannels written by Fabio Mammano and has been published by Springer Nature this book supported file pdf, txt, epub, kindle and other format this book has been release on 2024-04-05 with Science categories.
This volume explores the latest developments in the study of connexin hemichannels (HCs). The chapters in this book cover topics such as purification, reconstitution, and functional analysis of connexin HCs; spatial and temporal localization of connexins in cells using confocal microscopy; evaluation of connexin HC activity in vivo; generation of connexin-expressing stable cell pools; and automated assessment of cutaneous pathology in a mouse model of HC dysfunction. 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. Cutting-edge and comprehensive, Connexin Hemichannels: Methods and Protocols is a valuable resource that discusses the importance of HCs as drug targets, and how studying them is essential to identifying newcompounds capable of modulating HC function selectively and specifically.
Calcium Binding Protein Protocols
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Author : Hans J. Vogel
language : en
Publisher: Springer Science & Business Media
Release Date : 2008-02-04
Calcium Binding Protein Protocols written by Hans J. Vogel and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2008-02-04 with Science categories.
Calcium plays an important role in a wide variety of biological processes. This divalent metal ion can bind to a large number of proteins; by doing so it modifies their biological activity or their stability. Because of its distinct che- cal properties calcium is uniquely suited to act as an on–off switch or as a light dimmer of biological activities. The two books entitled Calcium-Binding Protein Protocols (Volumes I and II) focus on modern experimental analyses and methodologies for the study of calcium-binding proteins. Both extracel- lar and intracellular calcium-binding proteins are discussed in detail. H- ever, proteins involved in calcium handling (e. g. , calcium pumps and calcium channels), fall outside of the scope of these two volumes. Also, calcium-bi- ing proteins involved in bone deposition will not be discussed, as this specific topic has been addressed previously. The focus of these two books is on studies of the calcium-binding proteins and their behavior in vitro and in vivo. The primary emphasis is on protein chemistry and biophysical methods. Many of the methods described will also be applicable to proteins that do not bind calcium. Calcium-Binding Protein Protocols is divided into three main sections. The section entitled Introduction and Reviews provides information on the role of calcium in intracellular secondary messenger activation mechanisms. Mo- over, unique aspects of calcium chemistry and the utilization of calcium in dairy proteins, as well as calcium-binding proteins involved in blood clotting, are addressed.
Oxidative Stress Biomarkers And Antioxidant Protocols
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Author : Donald Armstrong
language : en
Publisher: Springer Science & Business Media
Release Date : 2008-02-04
Oxidative Stress Biomarkers And Antioxidant Protocols written by Donald Armstrong and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2008-02-04 with Science categories.
The first protocols book, Free Radical and Antioxidant Protocols (1) was published in late 1998. Sections were divided into three parts, covering selected biochemical techniques for measuring oxidative stress, antioxidant (AOX) activity, and combined applications. In choosing the 40 methods to be included in that book, I realized there were considerably more of equal value than that which we could have presented in a single volume. To produce a comprehensive resource, this book and a third are being compiled to expand coverage of the field. A summary of papers (2) published on this important subject emphasizes the continuing rapid growth in oxidative stress investigations relating to our understanding of biochemical reactions, their relevance to pathophysiological mechanisms, how disease may arise, and how therapeutic intervention may be achieved(3). Although there is some overlap between the categories, the ana- sis shown below illustrates where current studies are concentrated and are almost evenly distributed between free radicals and AOX. Over the last 4 yr, there has been a 55% increase in the number of papers published in the area.
Gtpase Protocols
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Author : Ed Manser
language : en
Publisher: Springer Science & Business Media
Release Date : 2008-02-03
Gtpase Protocols written by Ed Manser and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2008-02-03 with Science categories.
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.
Epithelial Cell Culture Protocols
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Author : Clare Wise
language : en
Publisher: Springer Science & Business Media
Release Date : 2008-02-05
Epithelial Cell Culture Protocols written by Clare Wise and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2008-02-05 with Medical categories.
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.
Genomics Protocols
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Author : Michael P. Starkey
language : en
Publisher: Springer Science & Business Media
Release Date : 2008-02-03
Genomics Protocols written by Michael P. Starkey and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2008-02-03 with Medical categories.
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).
Transgenesis Techniques
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Author : Alan R. Clarke
language : en
Publisher: Springer Science & Business Media
Release Date : 2008-02-05
Transgenesis Techniques written by Alan R. Clarke and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2008-02-05 with Science categories.
The past decade has witnessed a spectacular explosion in both the devel- ment and use of transgenic technologies. Not only have these been used to aid our fundamental understanding of biologic mechanisms, but they have also faci- tated the development of a range of disease models that are now truly beginning to impact upon our approach to human disease. Some of the most exciting model systems relate to neurodegenerative disease and cancer, where the availability of appropriate models is at last allowing radically new therapies to be developed and tested. This latter point is of particular significance given the current concerns of the wider public over both the use of animal models and the merits of using genetically modified organisms. Arguably, advances of the greatest significance have been made using mammalian systems—driven by the advent of embryonic stem-cell–based strategies and, more recently, by cloning through nuclear transfer. For this reason, this new edition of Transgenesis Techniques focuses much more heavily on manipulation of the mammalian genome, both in the general discussions and in the provision of specific protocols.
Pcr Cloning Protocols
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Author : Bing-Yuan Chen
language : en
Publisher: Springer Science & Business Media
Release Date : 2008-02-05
Pcr Cloning Protocols written by Bing-Yuan Chen and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2008-02-05 with Science categories.
PCR Cloning Protocols, Second Edition, updates and expands Bruce White's best-selling PCR Cloning Protocols (1997) with the newest procedures for DNA cloning and mutagenesis. Here the researcher will find readily reproducible methods for all the major aspects of PCR use, including PCR optimization, computer programs for PCR primer design and analysis, and novel variations for cloning genes of special characteristics or origin, with emphasis on long distance PCR and GC-rich template amplification. Also included are both conventional and novel enzyme-free and restriction site-free procedures to clone PCR products into a range of vectors, as well as state-of-the-art protocols to facilitate DNA mutagenesis and recombination, and to clone the challenging uncharacterized DNA flanking a known DNA fragment.
In Vitro Mutagenesis Protocols
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Author : Jeff Braman
language : en
Publisher: Springer Science & Business Media
Release Date : 2008-02-05
In Vitro Mutagenesis Protocols written by Jeff Braman and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2008-02-05 with Science categories.
Hands-on researchers with proven track records describe in stepwise fashion their advanced mutagenesis techniques. The contributors focus on improvements to conventional site-directed mutagenesis, including a chapter on chemical site-directed mutagenesis, PCR-based mutagenesis and the modifications that allow high throughput mutagenesis experiments, and mutagenesis based on gene disruption (both in vitro- and in situ-based). Additional methods are provided for in vitro gene evolution; for gene disruption based on recombination, transposon, and casette mutagenesis; and for facilitating the introduction of multiple mutations. Time-tested and highly practical, the protocols in In Vitro Mutagenesis Protocols, 2nd Edition offer today's molecular biologists reliable and powerful techniques with which to illuminate the proteome.