Immunocytochemical Methods and Protocols

Immunocytochemical Methods and Protocols
Author: Constance Oliver
Publisher: Humana Press
Total Pages: 416
Release: 2012-04-06
Genre: Medical
ISBN: 9781617796821

Antibodies tagged with fuorescent markers have been used in histochemistry for over 50 years. Although early applications were focused on the detection of microbial antigens in tissues, the use of immunocytochemical methods now has spread to include the det- tion of a wide array of antigens including proteins, carbohydrates, and lipids from virtually any organism. Today, immunohistochemistry is widely used to identify, in situ, various components of cells and tissues in both normal and pathological conditions. The method gains its strength from the extremely sensitive interaction of a specifc antibody with its antigen. For some scientifc areas, books have been published on applications of immu- cytochemical techniques specifc to that area. What distinguished Immunocytochemical Methods and Protocols from earlier books when it was frst published was its broad appeal to investigators across all disciplines, including those in both research and clinical settings. The methods and protocols p- sented in the frst edition were designed to be general in their application; the accompa- ing “Notes” provided the reader with invaluable assistance in adapting or troubleshooting the protocols. These strengths continued to hold true for the second edition and again for the third edition. Since the publication of the frst edition, the application of immuno- tochemical techniques in the clinical laboratory has continued to rise and this third edition provides methods that are applicable to basic research as well as to the clinical laboratory.

Immunocytochemical Methods and Protocols

Immunocytochemical Methods and Protocols
Author: Lorette C. Javois
Publisher: Springer Science & Business Media
Total Pages: 928
Release: 2008-02-02
Genre: Science
ISBN: 1592592139

Lorette Javois' timely new 2nd edition revises and updates her widely acclaimed collection of step-by-step immunocytochemical methods, one that is now used in many biological and biomedical research programs. The methods are designed for researchers and clinicians who wish to visualize molecules in plant or animal embryos, tissue sections, cells, or organelles. In addition to cutting-edge protocols for purifying and preparing antibodies, light microscopic analysis, confocal microscopy, FACS, and electron microscopy, this revised edition contains many new methods for applying immunocytochemical techniques in the clinical laboratory and in combination with in situ hybridization.

Electron Microscopy Methods and Protocols

Electron Microscopy Methods and Protocols
Author: M. A. Nasser Hajibagheri
Publisher: Springer Science & Business Media
Total Pages: 292
Release: 2008-02-02
Genre: Science
ISBN: 1592592015

Electron Microscopy Methods and Protocols is designed for the established researcher as a manual for extending knowledge of the field. It is also for the newcomer who wishes to move into the field. A wide range of applications for the examination of cells, tissues, biological macromolecules, molecular structures, and their interactions are discussed. We have tried to gather together methods that we consider to be those most generally appli- ble to current research in both cell and molecular biology. Each chapter c- tains a set of related practical protocols with examples provided by experts who have first-hand knowledge of the techniques they describe. The individual chapters are grouped according to similarities in their specimen preparation and methodology. Methods are presented in detail, in a step-by-step fashion, using reproducible protocols the authors have personally checked. During the last decade, the scientific literature describing the use of colloidal gold as an immunocytochemical marker has increased at an ex- nential rate, and this trend is expected to continue. We have included a large number of variations on the immunogold labeling technique. In both the ne- tive staining and cryo chapters, authors emphasize the “immunological app- cations” in order to correlate as fully as possible with the emphasis on immunogold labeling in the other chapters. Electron Microscopy Methods and Protocols commences with the routine preparation of biological material for classical transmission electron microscopy involving tissue fixation, embedding, and sectioning (Chap. 1).

Nuclease Methods and Protocols

Nuclease Methods and Protocols
Author: Catherine H. Schein
Publisher: Springer Science & Business Media
Total Pages: 521
Release: 2008-02-03
Genre: Science
ISBN: 1592592333

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.

Human Pluripotent Stem Cells

Human Pluripotent Stem Cells
Author: Philip H. Schwartz
Publisher: Humana Press
Total Pages: 468
Release: 2011-08-09
Genre: Science
ISBN: 9781617792007

Almost daily, new technologies are being presented that move the field of human pluripotent stem cell research towards a future that may yield highly-effective, personalized medical treatments. Three enabling technologies at hand for human PSCs are 1) directed reprogramming of somatic cells, which eliminate many of the ethical issues associated with the derivation and use of human PSCs, increase genetic diversity of the available human PSC lines, and give rise to better in vitro human disease models; 2) the discovery that a Rho-associated protein Kinase (ROCK) inhibitor allows for efficient single cell passaging and cryopreservation, increasing the efficiency and reliability of hPSC culture; and 3) defined, animal-component-free media, which lay the groundwork for simplified scale-up for therapeutic applications, differentiation protocols, and toxicology screens. The aforementioned technologies can be found in Human Pluripotent Stem Cells: Methods and Protocols, a compilation of 33 detailed protocols in six categories of PSC research that cover laboratory essentials and the derivation of new PSC lines, including induced PSC lines, as well as their growth, maintenance, characterization, genetic manipulation, and differentiation. Written in the successful Methods in Molecular BiologyTM series format, 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 accessible, Human Pluripotent Stem Cells: Methods and Protocols serves as an ideal guide to scientists conducting their own pluripotent cell research programs and makes great strides towards furthering human knowledge and, ultimately, improving the human condition.

Immunocytochemical Methods and Protocols

Immunocytochemical Methods and Protocols
Author: Lorette C. Javois
Publisher: Springer Science & Business Media
Total Pages: 435
Release: 1994
Genre: Medical
ISBN: 9780896032859

Annotation A collection of step-by-step immunocytochemical methods that can be used in many biological and biomedical disciplines in both research and clinical settings. Among the especially valuable protocols presented are those geared to preparing antibodies for staining procedures, light microscopic analysis, FACS, electron microscopy, and confocal microscopy. Each protocol includes troubleshooting tips and alternative procedures, as well as explanations of why certain steps are necessary, and comprehensive lists of reagents and suppliers. Plastic comb-binding. Annotation c. by Book News, Inc., Portland, Or.

Immunohistochemistry and Immunocytochemistry

Immunohistochemistry and Immunocytochemistry
Author: Luis Del Valle
Publisher:
Total Pages: 286
Release: 2022
Genre: Electronic books
ISBN: 9781071619483

This volume provides a comprehensive reference guide for researchers to study the applications of labeled antibodies. Chapters guide reader through the the and practice of immunohistochemistry, immunocytochemistry and immunofluorescence techniques. 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 useful tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Immunohistochemistry and Immunofluorescence: Methods and Protocols aims to be a useful practical guide to scientists to help further their study in this field.

Immunohistochemistry: Basics and Methods

Immunohistochemistry: Basics and Methods
Author: Igor B. Buchwalow
Publisher: Springer Science & Business Media
Total Pages: 153
Release: 2010-01-26
Genre: Medical
ISBN: 3642046096

This concise yet comprehensive guide to the methods and protocols of immunohistochemistry covers established techniques and current developments in the field such as the use of epitope tags, multiple immunolabeling and diagnostic immunohistochemistry.

Adipose Tissue Protocols

Adipose Tissue Protocols
Author: Gérard Ailhaud
Publisher: Springer Science & Business Media
Total Pages: 332
Release: 2008-02-03
Genre: Science
ISBN: 1592592317

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.

Antibody Engineering Protocols

Antibody Engineering Protocols
Author: Sudhir Paul
Publisher: Springer Science & Business Media
Total Pages: 445
Release: 2008-02-02
Genre: Medical
ISBN: 1592595383

This comprehensive collection of recently developed methods for producing new antibody reagents by immunization and recombinant DNA techniques contains ready-to-use protocols that illuminate current areas of research on antibody structure, functions, and applications. The methods can be applied in basic immunological studies involving antibody specificity, catalysis, and evolution, and in the isolation of rare antibodies by phage display technology and the engineering of new antibodies by mutagenesis. They offer insight into new ways of developing clinically useful antibody reagents. Antibody Engineering Protocols constitutes a single-source volume for laboratory investigators who want to minimize extensive literature and methodology searches and to work productively in their fields with reproducible step-by-step protocols.