Proteoglycan Protocols

Proteoglycan Protocols

Author: Renato V. Iozzo

Publisher: Springer Science & Business Media

Published: 2008-02-02

Total Pages: 547

ISBN-13: 1592592090

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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.


Glycomics

Glycomics

Author: Nicolle H. Packer

Publisher: Humana Press

Published: 2009-07-16

Total Pages: 390

ISBN-13: 9781607612407

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Due to the significant contributions of carbohydrates to the functional diversity of the cell, the challenging study of the glycome has expanded beyond the research of carbohydrate experts and into the wider scope of the life sciences. To aid all scientists now delving into this vital subject area, Glycomics: Methods and Protocols collects a compendium of detailed laboratory protocols reflecting the increasing availability of sample preparation, chromatographic, electrophoretic, mass spectrometric, and bioinformatic tools specifically designed for the analysis of glycosylation. Leading researchers in the field address subjects such as glycoprotein and proteoglycan analysis, glycosylation structure determination, as well as various approaches to investigate the interaction between glycans and a variety of carbohydrate-recognizing proteins in order to aid exploration into the functional significance of the oligosaccharides. Written in the highly successful Methods in Molecular BiologyTM series format, the 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 cutting-edge, Glycomics: Methods and Protocols serves as a valuable guide for experimenters facing the challenges of glycan analysis in hope of providing further insights into the biology of cell-cell communication and interaction.


Cell-Penetrating Peptides

Cell-Penetrating Peptides

Author: Ülo Langel

Publisher: Humana

Published: 2015-07-24

Total Pages: 0

ISBN-13: 9781493928057

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Divided into three parts this volume summarizes the most important areas of Cell-Penetrating Peptides (CPP) research . Part one briefly presents the historical background of CPP studies and the classifications of the available CPPs, and then summarizes the approaches for prediction of novel CPPs. Part two mainly describes the methods for studies of “naked” CPPs, that is, CPPs without conjugated cargos. Last but not least part three presents a representative and brief summary of functionality issues of CPPs, both in vitro and in vivo. As a volume in the highly successful Methods in Molecular Biology series, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls. Concise and easy-to-use, Cell-Penetrating Peptides: Methods and Protocols, Second Edition hopes to raise relevant questions for further development.


Glycosaminoglycans and Proteoglycans

Glycosaminoglycans and Proteoglycans

Author: 978-3-03842-836-7

Publisher: MDPI

Published: 2018-07-04

Total Pages: 247

ISBN-13: 3038428353

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This book is a printed edition of the Special Issue "Glycosaminoglycans and Proteoglycans" that was published in Pharmaceuticals


Essentials of Glycobiology

Essentials of Glycobiology

Author: Ajit Varki

Publisher: CSHL Press

Published: 1999

Total Pages: 694

ISBN-13: 9780879696818

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Sugar chains (glycans) are often attached to proteins and lipids and have multiple roles in the organization and function of all organisms. "Essentials of Glycobiology" describes their biogenesis and function and offers a useful gateway to the understanding of glycans.


Adipose Tissue Protocols

Adipose Tissue Protocols

Author: Gérard Ailhaud

Publisher: Springer Science & Business Media

Published: 2008-02-03

Total Pages: 332

ISBN-13: 1592592317

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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.


In Vitro Mutagenesis Protocols

In Vitro Mutagenesis Protocols

Author: Jeff Braman

Publisher: Springer Science & Business Media

Published: 2008-02-05

Total Pages: 284

ISBN-13: 1592591949

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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.


DNA'Protein Interactions

DNA'Protein Interactions

Author: Tom Moss

Publisher: Springer Science & Business Media

Published: 2008-02-02

Total Pages: 633

ISBN-13: 1592592082

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Dr. Tom Moss assembles the new standard collection of cutting-edge techniques to identify key protein-DNA interactions and define their components, their manner of interaction, and their manner of function, both in the cell and in the test tube. The techniques span a wide range, from factor identification to atomic detail, and include multiple DNA footprinting analyses, including in vivo strategies, gel shift (EMSA) optimization, SELEX, surface plasmon resonance, site-specific DNA-protein crosslinking, and UV laser crosslinking. Comprehensive and broad ranging, DNA-Protein Interactions: Principles and Protocols, 2nd Edition, offers a stellar array of over 100 up-to-date and readily reproducible techniques that biochemists and molecular, cellular, and developmental biologists can use successfully today to understand DNA-protein interactions.


Two-Hybrid Systems

Two-Hybrid Systems

Author: Paul N. MacDonald

Publisher: Springer Science & Business Media

Published: 2008-02-03

Total Pages: 337

ISBN-13: 1592592104

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The yeast two-hybrid system is one of the most widely used and productive techniques available for investigating the macromolecular interactions that affect virtually all biological processes. In Two-Hybrid Systems: Methods and Protocols, Paul N. MacDonald has assembled a collection of these powerful molecular tools for examining and characterizing protein-protein, protein-DNA, and protein-RNA interactions. The techniques range from the most basic (introducing plasmids into yeasts, interaction assays, and recovering the plasmids from yeast) to the most advanced alternative strategies (involving one-hybrid, split two-hybrid, three-hybrid, membrane recruitment systems, and mammalian systems). Methods are also provided for dealing with the well-known problems of artifacts and false positives and for identifying the interacting partners in important biological systems, including the Smad and nuclear receptor pathways. To ensure ready reproducibility and robust results, each technique is described in step-by-step detail by researchers who employ it regularly. Comprehensive and highly practical, Two-Hybrid Systems: Methods and Protocols not only reveals how the great variety of plasmid vectors and approaches may be optimally deployed, but also quickly empowers novices to establish two-hybrid systems in their laboratories, and experienced researchers to expand their repertoire of techniques.


DNA Arrays

DNA Arrays

Author: Jang B. Rampal

Publisher: Springer Science & Business Media

Published: 2008-02-02

Total Pages: 268

ISBN-13: 1592592341

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Microarray technology provides a highly sensitive and precise te- nique for obtaining information from biological samples, with the added advantage that it can handle a large number of samples simultaneously that may be analyzed rapidly. Researchers are applying microarray technology to understand gene expression, mutation analysis, and the sequencing of genes. Although this technology has been experimental, and thus has been through feasibility studies, it has just recently entered into widespread use for advanced research. The purpose of DNA Arrays: Methods and Protocols is to provide instruction in designing and constructing DNA arrays, as well as hybridizing them with biological samples for analysis. An additional purpose is to p- vide the reader with a broad description of DNA-based array technology and its potential applications. This volume also covers the history of DNA arrays—from their conception to their ready off-the-shelf availability—for readers who are new to array technology as well as those who are well versed in this field. Stepwise, detailed experimental procedures are described for constructing DNA arrays, including the choice of solid support, attachment methods, and the general conditions for hybridization. With microarray technology, ordered arrays of oligonucleotides or other DNA sequences are attached or printed to the solid support using au- mated methods for array synthesis. Probe sequences are selected in such a way that they have the appropriate sequence length, site of mutation, and T .