Calpain Methods And Protocols

Author : John S. Elce
Publisher : Springer Science & Business Media
Page : 340 pages
File Size : 45,9 Mb
Release : 2008-02-05
Category : Medical
ISBN : 9781592590506

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Calpain Methods and Protocols by John S. Elce Pdf

The purpose of Calpain Methods and Protocols is quite straightf- ward: it is to present the actual experimental methods used in many different laboratories for the study of calpain. It will provide the vital experimental detail, and the discussion of possible pitfalls, for which the standard journals no longer provide space. This will make it as easy as possible for investi- tors interested in calpain to adopt established methods without repeating old mistakes, and to adapt and apply these methods in novel approaches to the many outstanding calpain questions. These questions range from purely biochemical problems of protein structure and enzyme regulation at the molecular level, through large areas of cell biology, to applied and clinical aspects of calpain function in human d- ease. Within this panoply of topics, a wide range of investigators will find many fascinating and as yet unanswered questions about calpain. Calpain Methods and Protocols will provide instant access to many essential te- niques, while saving them the time and effort involved in developing a new method. In addition to questions relating to the normal physiological roles of the calpains, there is considerable evidence that inappropriate calpain activity may have pathological effects in many tissues, for example, following ischemia. This provides a major stimulus for the development of specific calpain inhi- tors for therapeutic purposes, and for the development of methods to evaluate such inhibitors.

Author : John M. Walker
Publisher : Unknown
Page : 128 pages
File Size : 54,5 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: Calpain methods and protocols by John M. Walker Pdf

Author : John S. Elce
Publisher : Humana Press
Page : 342 pages
File Size : 53,9 Mb
Release : 2000-02-21
Category : Medical
ISBN : 0896036324

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Calpain Methods and Protocols by John S. Elce Pdf

The purpose of Calpain Methods and Protocols is quite straightf- ward: it is to present the actual experimental methods used in many different laboratories for the study of calpain. It will provide the vital experimental detail, and the discussion of possible pitfalls, for which the standard journals no longer provide space. This will make it as easy as possible for investi- tors interested in calpain to adopt established methods without repeating old mistakes, and to adapt and apply these methods in novel approaches to the many outstanding calpain questions. These questions range from purely biochemical problems of protein structure and enzyme regulation at the molecular level, through large areas of cell biology, to applied and clinical aspects of calpain function in human d- ease. Within this panoply of topics, a wide range of investigators will find many fascinating and as yet unanswered questions about calpain. Calpain Methods and Protocols will provide instant access to many essential te- niques, while saving them the time and effort involved in developing a new method. In addition to questions relating to the normal physiological roles of the calpains, there is considerable evidence that inappropriate calpain activity may have pathological effects in many tissues, for example, following ischemia. This provides a major stimulus for the development of specific calpain inhi- tors for therapeutic purposes, and for the development of methods to evaluate such inhibitors.

Author : Roberto Bruzzone,Christian Giaume
Publisher : Springer Science & Business Media
Page : 495 pages
File Size : 44,9 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.

Author : Catherine H. Schein
Publisher : Springer Science & Business Media
Page : 521 pages
File Size : 54,5 Mb
Release : 2008-02-03
Category : Science
ISBN : 9781592592333

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Nuclease Methods and Protocols by Catherine H. Schein Pdf

Nucleases, enzymes that restructure or degrade nucleic acid polymers, are vital to the control of every area of metabolism. They range from “housekeeping” enzymes with broad substrate ranges to extremely specific tools (1). Many types of nucleases are used in lab protocols, and their commercial and clinical uses are expanding. The purpose of Nuclease Methods and Protocols is to introduce the reader to some we- characterized protein nucleases, and the methods used to determine their activity, structure, interaction with other molecules, and physiological role. Each chapter begins with a mini-review on a specific nuclease or a nuclease-related theme. Although many chapters cover several topics, they were arbitrarily divided into five parts: Part I, “Characterizing Nuclease Activity,” includes protocols and assays to determine general (processive, distributive) or specific mechanisms. Methods to assay nuclease products, identify cloned nucleases, and determine their physiological role are also included here. Part II, “Inhibitors and Activators of Nucleases,” summarizes assays for measuring the effects of other proteins and small molecules. Many of these inhibitors have clinical relevance. Part III, “Relating Nuclease Structure and Function,” provides an overview of methods to determine or model the 3-D structure of nucleases and their complexes with substrates and inhibitors. A 3-D structure can greatly aid the rational design of nucleases and inhibitors for specific purposes. Part IV, “Nucleases in the Clinic,” summarizes assays and protocols suitable for use with t- sues and for nuclease based therapeutics.

Author : Tom Moss
Publisher : Springer Science & Business Media
Page : 633 pages
File Size : 47,9 Mb
Release : 2008-02-02
Category : Science
ISBN : 9781592592081

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DNA'Protein Interactions by Tom Moss Pdf

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.

Author : Gérard Ailhaud
Publisher : Springer Science & Business Media
Page : 332 pages
File Size : 55,6 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 : Martin J. Tymms,Ismail Kola
Publisher : Springer Science & Business Media
Page : 434 pages
File Size : 44,8 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 : Amanda E.I. Proudfoot,Timothy N.C. Wells,Christine Power
Publisher : Springer Science & Business Media
Page : 349 pages
File Size : 51,9 Mb
Release : 2008-02-05
Category : Medical
ISBN : 9781592590582

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Chemokine Protocols by Amanda E.I. Proudfoot,Timothy N.C. Wells,Christine Power Pdf

The chemokines family of small proteins are involved in numerous b- logical processes ranging from hematopoiesis, angiogenesis, and basal l- kocyte trafficking to the extravasation and tissue infiltration of leukocytes in response to inflammatory agents, tissue damage, and bacterial or viral infection. Chemokines exert their effects through a family of seven G-protein coupled transmembrane receptors. Worldwide interest in the chemokine field surged dramatically early in 1996, with the finding that certain chemokine receptors were the elusive coreceptors, required along with CD4, for HIV infection. Today, though over 40 human chemokines have been described, the n- ber of chemokine receptors lags behind—only 17 human chemokine receptors have been identified so far. What has emerged over the years is that most chemokine receptors bind several distinct ligands, and indeed the majority of chemokines are able to bind to multiple chemokine receptors, explaining to some extent the apparent disparity in the numbers of chemokines and rec- tors. Yet in spite of the apparent redundancy in chemokine/chemokine rec- tor interactions, it is clear that in vivo, spatial, temporal, and indeed cell- and tissue-specific expression of both chemokines and their receptors are imp- tant factors in determining the precise nature of cellular infiltrates in phy- ological and pathological processes.

Author : Robert A. Rush
Publisher : Springer Science & Business Media
Page : 272 pages
File Size : 40,9 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 : Philip H. Howe
Publisher : Springer Science & Business Media
Page : 176 pages
File Size : 50,9 Mb
Release : 2008-02-05
Category : Science
ISBN : 9781592590537

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Transforming Growth Factor-Beta Protocols by Philip H. Howe Pdf

Philip H. Howe and a group of well-versed experimentalists present the first major volume a collection of indispensable classic and cutting-edge TGFß assays. Described in great detail to ensure robust and successful results, these readily reproducible techniques range from the growth inhibition assay for TGFß to methods for monitoring its interactions with the mediating proteins. Extensive notes discuss potential pitfalls and provide tips on how to avoid failures, and throughout, emphasis is given to detailing those technical steps critical for experimental success that are often omitted in the primary literature. Concise and highly practical, Transforming Growth Factor-Beta Protocols provides today's molecular and cell biologists-both expert and novice-with time-tested methods for the identification and analysis of the signal transduction pathways by which TGFß induces and modulates physiological behavior.

Author : Ian M. Clark
Publisher : Springer Science & Business Media
Page : 547 pages
File Size : 45,5 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 : Mary W. Trucksess,Albert E. Pohland
Publisher : Springer Science & Business Media
Page : 245 pages
File Size : 47,6 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 : Catherine Cooper
Publisher : Springer Science & Business Media
Page : 268 pages
File Size : 42,9 Mb
Release : 2008-02-05
Category : Science
ISBN : 9781592590476

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Amino Acid Analysis Protocols by Catherine Cooper Pdf

A collection of classic and cutting-edge techniques of high utility in answering specific biological questions about amino acids. Common methods include those based on HPLC or gas chromatography separation and analysis after precolumn derivatization. New techniques based on capillary electrophoresis separation, high-performance anion exchange chromatography, and mass spectrometry are also presented. Each method is described in step-by-step detail to ensure successful experimental results and emphasizes sample preparation, particularly the collection and storage of bodily fluids. Up-to-date and highly practical, Amino Acid Analysis Protocols offers analytical and clinical chemists, as well as a broad range of biological and biomedical investigators, a rich compendium of laboratory tools for the productive analysis of both common and uncommon amino acids.

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.