Embryonic Stem Cell Protocols


Book Description

Now in two volumes, this completely updated and expanded edition of Embryonic Stem Cells: Methods and Protocols provides a diverse collection of readily reproducible cellular and molecular protocols for the manipulation of nonhuman embryonic stem cells. Volume one, Embryonic Stem Cell Protocols: Isolation and Characterization, Second Edition, provides a diverse collection of readily reproducible cellular and molecular protocols for the isolation, maintenance, and characterization of embryonic stem cells. The second volume, Embryonic Stem Cell Protocols: Differentiation Models, Second Edition, covers state-of-the-art methods for deriving many types of differentiating cells from ES cells. Together, the two volumes illuminate for both novices and experts our current understanding of the biology of embryonic stem cells and their utility in normal tissue homeostasis and regenerative medicine applications.




Developmental Biology Protocols


Book Description

Developmental biology is one of the most exciting and fast-growing fields today. In part, this is so because the subject matter deals with the innately fascinating biological events—changes in form, structure, and function of the org- ism. The other reason for much of the excitement in developmental biology is that the field has truly become the unifying melting pot of biology, and provides a framework that integrates anatomy, physiology, genetics, biochemistry, and cellular and mole- lar biology, as well as evolutionary biology. No longer is the study of embryonic development merely “embryology.” In fact, development biology has produced - portant paradigms for both basic and clinical biomedical sciences alike. Although modern developmental biology has its roots in “experimental emb- ology” and the even more classical “chemical embryology,” the recent explosive and remarkable advances in developmental biology are critically linked to the advent of the “cellular and molecular biology revolution.” The impressive arsenal of expe- mental and analytical tools derived from cell and molecular biology, which promise to continue to expand, together with the exponentially developing sophistication in fu- tional imaging and information technologies, guarantee that the study of the devel- ing embryo will contribute one of the most captivating areas of biological research in the next millennium.







Developmental Biology Protocols


Book Description

Developmental biology is one of the most exciting and fast-growing fields today. In part, this is so because the subject matter deals with the innately fascinating biological events—changes in form, structure, and function of the org- ism. The other reason for much of the excitement in developmental biology is that the field has truly become the unifying melting pot of biology, and provides a framework that integrates anatomy, physiology, genetics, biochemistry, and cellular and mole- lar biology, as well as evolutionary biology. No longer is the study of embryonic development merely “embryology.” In fact, development biology has produced - portant paradigms for both basic and clinical biomedical sciences alike. Although modern developmental biology has its roots in “experimental emb- ology” and the even more classical “chemical embryology,” the recent explosive and remarkable advances in developmental biology are critically linked to the advent of the “cellular and molecular biology revolution.” The impressive arsenal of expe- mental and analytical tools derived from cell and molecular biology, which promise to continue to expand, together with the exponentially developing sophistication in fu- tional imaging and information technologies, guarantee that the study of the devel- ing embryo will contribute one of the most captivating areas of biological research in the next millennium.




Plant Developmental Biology


Book Description

Plants come in myriads of shapes and colors, and the beauty of plants has fascinated mankind for thousands of years. Long before Mendel discovered the laws of heritab- ity and Darwin developed his theory on evolution, the affection for ornamental plants led people to select alleles that establish novel plant forms. Today, plant developmental biology tries to discover the mechanisms that control the establishment of specialized cell types, tissues, and organs from the fertilized egg during a plant’s life. Although the underlying processes of cell proliferation and differentiation are similar in plants and a- mals, plants are different because their development is usually open, and its outcome is not the faithful repetition of a general plan but is strongly in?uenced by environm- tal conditions. In the last few decades, plant developmental biology has pinpointed a large number of developmental regulators and their interactions and the mechanisms that govern plant development start to emerge. In part, this progress was enabled by the advance of powerful molecular tools for a few model species, most importantly Arabidopsis. This volume of the Methods in Molecular Biology series provides a collection of protocols for many of the common experimental approaches in plant developmental bi- ogy. All chapters are written in the same format as that used in the Methods in Molecular TM Biology series. Each chapter opens with a description of the basic theory behind the method being described.




Apoptosis Methods and Protocols


Book Description

The most fundamental question facing each and every cell within an org- ism is to survive or to die. Cell death is required for normal function; some estimates suggest that as many as one million cells undergo cell death every second in the adult human body. Almost all cells undergoing physiological, or programmed, cell death, independent of cell type, manifest a stereotypic p- tern of morphological changes termed apoptosis. Typically, apoptotic cells d- play shrinkage, membrane blebbing, chromatin condensation, and nuclear fragmentation. The integrity of the cell membrane is not lost during apoptosis and so avoids eliciting the inflammatory response that would have been caused by the spillage of the cell’s contents. This is quite in contrast to the loss of cell contents typical of necrosis. The caspases, the family of intracellular cysteine proteases associated with apoptosis, are responsible for the stereotypical m- phological changes. Caspases cleave various substrate proteins that act on DNA fragmentation, nuclear envelope integrity, the cytoskeleton, and cell volume regulation. Apoptotic cells are cleared in vivo by the process of phagocytosis, in which specific “phagocytes” move to the site of apoptosis, engulf the dying cells and digest them. Apoptosis has a central role in many physiological processes, for example, in the immune system. Autoreactive cells are deleted via apoptosis to prevent autoimmunity. At the end of an immune response, activated lymphocytes are removed to maintain homeostasis within the immune system.




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.




The Kidney


Book Description

Organogenesis of the kidney has been intensely studied for over a century. In recent years advances in molecular techniques have not only made great inroads into exploring the genetic regulation of this complex process but also began to unravel the molecular basis of many forms of congenital kidney disease. This book is a comprehensive study on these findings and the only book available with such in depth coverage of the kidney. - Hundreds of color figures depicting key events in all aspects of kidney development - Full coverage of the genetic and cellular basis of kidney development - Analysis of the genetic basis of the major congenital kidney diseases




DNA Topoisomerase Protocols


Book Description

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.




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.