Search results for: transgenic-mouse-methods-and-protocols

Transgenic Mouse

Author : Marten H. Hofker
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A collection of readily reproducible methods for working with mice, and particularly for generating mouse models that will enable you to better understand gene function. Described in step-by-step detail by highly experienced investigators, these proven techniques include new methods for conditional, induced knockout, and transgenic mice, as well as for working with mice in such important research areas as immunology, cancer, and atherosclerosis. Such alternative strategies as random mutagenesis and viral gene transduction for studying gene function in the mouse are also presented. Use effective methods for conditional and inducible gene expression Review mouse models for a wide range of genetic aberrations associated with human disease.

Transgenic Mouse

Author : Melissa A. Larson
File Size : 40.87 MB
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This volume provides readers with a historical foundation in standard techniques and a comprehensive update on the latest methods used in making gene-modified mice. The chapters in this book cover topics such as pronuclear microinjection in one-cell embryos; embryo transfer surgery; nuclear transfer and cloning; blastocyst microinjection; and cryobanking and recovery of genetically modified mice. Importantly, there are chapters devoted to the latest application of CRISPR technology, as well as the establishment of induced pluripotent stem cells. 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. Comprehensive and authoritative, Transgenic Mouse: Methods and Protocols is a valuable resource for both novice and expert researchers who are interested in learning more about this evolving field.

Transgenic Mouse Methods and Protocols

Author : Marten H. Hofker
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Marten Hofker and Jan van Deursen have assembled a multidisciplinary collection of readily reproducible methods for working with mice, and particularlyfor generating mouse models that will enable us to better understand gene function. Described in step-by-step detail by highly experienced investigators, these proven techniques include new methods for conditional, induced knockout, and transgenic mice, as well as for working with mice in such important research areas as immunology, cancer, and atherosclerosis. Such alternative strategies as random mutagenesis and viral gene transduction for studying gene function in the mouse are also presented.

Protein Sequencing Protocols

Author : Bryan John Smith
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Featuring new and updated techniques for determining the sequence of proteins and peptides, this edition includes not only novel approaches to the validation of quality assurance methods, reflecting the current importance of biopharmaceuticals, but also offers a guide to analysis of protein sequence information via the powerful new tools of bioinformatics. Comprehensive and up-to-date, Protein Sequencing Protocols, Second Edition, provides for both novice and expert investigators alike a ready source of easy-to-follow protocols that simplify choosing the most appropriate method for protein sequence determination.

Superantigen Protocols

Author : Teresa Krakauer
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Leading researchers in the biological, chemical, and physical investigation of superantigens describe in step-by-step detail their best experimental techniques to assess the physical characteristics and biological effects of superantigens. Their protocols range from those for investigating the interactions of superantigens with cellular receptors to those for the analysis of their immunological and biological effects, including methods for using BIOcore to determine binding kinetics and establishing various lymphocyte cell culture systems. There are also accounts of such methods as the RNase protection assay, cytokine ELISA, FACS analysis, and cytokine production at the single cell level..

RT PCR Protocols

Author : Joe O'Connell
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A panel of highly regarded molecular biologists and clinical researchers describe in detail their most novel, useful, and interesting RT-PCR applications. Here the newcomer will find readily reproducible protocols for highly sensitive detection and quantification of gene expression, the in situ localization of gene expression in tissue, and the cloning of genes, as well as for analyzing T-cell clones and the differential expression of genes. For the expert seeking to extend the usefulness of RT-PCR, there are user-friendly applications that complement the latest technological advances, including laser-capture microdissection (LCM), real-time and quantitative PCR, microarray technology, cDNA cloning, and antibody engineering. Study disease pathogenesis with RT-PCR to design new therapeutic strategies Expand RT-PCR with antibody engineering, real-time PCR, and microarray technology.

Peptide Research Protocols

Author : Janet J. Maguire
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A panel of multidisciplinary experts describes in detail readily reproducible methods to investigate all aspects of the endothelin system from its synthesis and metabolism, to its function in health and disease. Theses methods use state-of-the-art molecular techniques to quantify the expression of mRNA for both endothelin receptors and the endothelin converting enzymes. They show how peptides, precursors, receptors, and synthetic enzymes can be localized and quantified in plasma, culture supernatants, tissue homogenate, and tissue sections using antibodies. Several in vivo protocols illustrate the role of the endothelin peptides in healthy human individuals and describe animal models that can be used to predict the therapeutic potential of cardiovascular drugs that manipulate endothelin synthesis or function.

Neutrophil Methods and Protocols

Author : Mark T. Quinn
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This book provides a concise set of protocols for assessing basic neutrophil functions, investigating specialized areas in neutrophil research, and completing step-by-step diagnostic assays of common neutrophil disorders. Each of the protocols is written by leading researchers in the field and includes hints for success, as well as guidance for troubleshooting. Scientists and clinicians will find this collection an invaluable aid.

Amyloid Proteins

Author : Einar M. Sigurdsson
File Size : 80.82 MB
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A proven collection of readily reproducible techniques for studying amyloid proteins and their involvement in the etiology, pathogenesis, diagnosis, and therapy of amyloid diseases. The contributors provide methods for the preparation of amyloid and its precursors (oligomers and protofibrils), in vitro assays and analytical techniques for their study, and cell culture models and assays for the production of amyloid proteins. Additional chapters present readily reproducible techniques for amyloid extraction from tissue, its detection in vitro and in vivo, as well as nontransgenic methods for developing amyloid mouse models. The protocols follow the successful Methods in Molecular Biology⢠series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.

Pain Research

Author : Z. David Luo
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Despite significant advances in molecular biology techniques and in our understanding of the physiology and the behavioral pharmacology of pain transduction, effective, target-specific therapeutic agents for chronic pain are still lacking. In Pain Research: Methods and Protocols, leading researchers who have first-hand experience describe, in step-by-step detail, diverse and novel techniques for dissecting the molecular mechanisms of pain transduction. These readily reproducible methods employ a variety of multidisciplinary approaches ranging from animal pain models and single neutron selection to in vitro single-cell mRNA amplification. The collection includes not only standard, cutting-edge methods, but also novel techniques only recently applied to pain research. The protocols follow the successful Methods in Molecular BiologyTM series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Versatile and easy-to-use, Pain Research: Methods and Protocols offers today's pain researchers in academic and pharmaceutical laboratories powerful tools to unravel the cellular and molecular complexity of pain transduction and set the stage for the next generation of pain medications.

Epidermal Cells

Author : Kursad Turksen
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Helps the reader to learn about the derivation, characterization, and utility of epidermal stem cells; follow step-by-step instructions that ensure successful results; understand the utility of epidermal cells in regenerative medicine applications; and apply reproducible methods to study epidermal precursors and mature epidermal cells.

Molecular Embryology

Author : Paul T. Sharpe
File Size : 77.70 MB
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In Molecular Embryology, expert investigators provide a comprehensive guide to the cutting-edge methods used today across the dramatically growing field of vertebrate molecular embryology. These powerful techniques take advantage of the most commonly used vertebrate experimental models: murine embryos for their genetics, chick embryos for in vivo manipulation, zebrafish for mutagenesis, amphibian embryos, and nonvertebrate chordates. The major techniques of experimental molecular biology and the particular advantages of each different species are emphasized. Detailed, easy-to-follow protocols, together with relevant background information and helpful tips, optimize the methods for success. Molecular Embryology brings together in one volume all the major techniques and common experimental species needed to study the mechanisms of biological development in vertebrates. Bound to become a standard reference in this field, the book makes it possible for experienced and novice researchers alike to move between embryos of diverse vertebrate classes as their project progresses, ensuring their ability to utilize the experimental advantages of different systems to address specific developmental questions.

Anticancer Research

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Cardiovascular Disease

Author : Qing Wang
File Size : 27.56 MB
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The two volumes of Cardiovascular Disease: Methods and Protocols provide comprehensive coverage of both basic and advanced approaches to the study and characterization of cardiovascular disease. In Volume 1: Genetics and Volume 2: Molecular Medicine, highly experienced cardiovascular researchers describe in detail the most important techniques in molecular medicine that are employed in genetic, molecular, cellular, structural, and physiological studies of cardiovascular disease.

Hypertension

Author : Jérôme P. Fennell
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Our understanding of the pathogenesis of hypertension and the development of new therapies for its management have each proven to be directly dependent on the availability and application of novel molecular biological methodologies. In Hypertension: Methods and Protocols, a panel of recognized experts from key institutions around the world describes in step-by-step detail many new and essential molecular techniques for cardiovascular hypertension research. The methods presented range widely from producing congenic, consomic, transgenic, and knockout models of hypertension to the gene transfer of specific genetic material using adenoviral and nonviral (polymers, liposomes, and antisense agents) vectors. Additional techniques described include single nucleotide polymorphism (SNP) genotyping, RNA interference, microarray analysis, pharmacogenetics, and pharmacogenomics for the genetic dissection of hypertension, as well as a practical method for deriving cardiomyocytes from embryonic stem cells that would serve as replacement cells for those damaged by hypertension or heart attack. A review of bioinformatic resources for pharmacogenomics describes the application of in silico strategies to identify nuclear matrix attachment regions. The protocols follow the successful Methods in Molecular Medicine series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. State-of-the-art and highly practical, Hypertension: Methods and Protocols offers both novice and experienced hypertension researchers an indispensable collection of readily reproducible techniques for successful research, work that has already dramatically improved the outlook for hypertensive patients, and promises much future success.

Gene Knockout Protocols

Author : Martin J. Tymms
File Size : 61.76 MB
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As the major task of sequencing the human genome nears completion and the full complement of human genes are catalogued, the task of understanding the normal biological functions of genes and how their alteration leads to diseased states becomes more imperative. In Gene Knockout Protocols, highly skilled investigators with extensive experience in gene targeting and mouse genetics describe their best techniques for the design of targeting constructs and for genetic phenotype analysis. These proven methods contain step-by-step instructions, as well as notes on pitfalls to avoid, and emphasize techniques that are relevant to researchers carrying out gene targeting work. These include embryo transplantation, in vitro embryonic stem cell differentiation, creation of aggregation chimeras, mouse pathology, embryo cryopreservation, and transplantation. Issues such as the use of existing mouse mutation resources and the influence of genetic background and epigenetic effects upon phenotype are also covered. State-of-the-art and highly practical, Gene Knockout Protocols not only constitutes an invaluable source of readily reproducible techniques for those just entering the field of gene targeting, but also a key reference for all genetic researchers today.

Placenta and Trophoblast

Author : Michael J. Soares
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Placenta Methods and Protocols represents a collection of detailed methodologies used in contemporary placental research. Techniques presented in the text range from comprehensive descriptions of experimental animal models, animal and human placental organ and cell culture systems, and morphological, biochemical, and molecular strategies for assessing trophoblast/placental growth, differentiation, and function.

Malaria Methods and Protocols

Author : Denise L. Doolan
File Size : 66.28 MB
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Despite considerable scientific and medical effort over the past decades, malaria remains the most important human parasitic disease. It is responsible for up to 3 million deaths and another 300-500 million new cases each year, and is becoming resistant to the current chemoprophylactic and chemotherapeutic agents. In Malaria Methods and Protocols, internationally respected scientists and clinicians describe in step-by-step detail their most useful conventional and cutting-edge techniques for the study of malaria. Areas covered include clinical and laboratory diagnosis and typing, animal models, molecular biology, immunology, cell biology, vaccinology, laboratory models, and field applications. Each readily reproducible protocol has been tested, standardized, and optimized for experimental success, and includes many laboratory notes on troubleshooting, avoiding pitfalls, and interpreting results. Several of the most widely used methods are either described here in detail for the first time or have been thoroughly updated since their original publication (e.g., in vitro culture of Plasmodium parasites and in vitro growth inhibition assay). State-of-the-art and highly practical, Malaria Methods and Protocols makes available to basic and applied researchers today's only comprehensive collection of essential laboratory methods for diagnosing malaria, characterizing the parasite, understanding the interaction between the human host and Plasmodium parasite, and developing effective preventive measures.

Interferon Methods and Protocols

Author : Daniel J. J. Carr
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A compendium of optimized methods to measure type I interferon efficacy as an antiproliferative or an antiviral agent. These cutting-edge techniques range from the simple to the highly complex and serve to illuminate the signaling cascades and the activation of enzymatic pathways prompted by interferon. State-of-the-art and highly practical, Interferon Methods and Protocols offers researchers powerful tools not only to ascertain the functions of IFN-stimulatory gene products, but also to identify additional molecular pathways that will clarify our understanding of the many biological events influenced by IFNs.

Nuclear Reprogramming

Author : Steve Pells
File Size : 48.50 MB
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This volume includes various methods of nuclear reprogramming, including nuclear transfer in several different species, both amphibian and mammalian, fusion achieved both by chemical treatment and by electrically shocking cells, and quantitative fusion and reprogramming in vitro treatment of cells with cell extracts. Also described are the different methods of monitoring nuclear programming after nuclear transfer, activation of telomerase as an ES-specific marker, observation of structural changes in the nucleus by both light and electron microscopy, and verification of stem cells' surface marker expression and differentiation potential.