Cell Cycle Control and Dysregulation Protocols: Cyclins, Cyclin-Dependent Kinases, and Other Factors

ISBN-13: 9781617372704 / Angielski / Miękka / 2010 / 200 str.

Antonio Giordano; Gaetano Romano

Opis

Cell Cycle Control and Dysregulation Protocols focuses on emerging methodologies for studying the cell cycle, kinases, and kinase inhibitors. It addresses the issue of gene expression in vivo and in vitro, the analysis of cyclin-dependent kinase inhibitors, protein degradation mediated by the proteosome, the analysis of the transformed cell phenotype, and innovative techniques to detect apoptosis. Because there are already many manuals and protocols available, along with commercial kits and reagents, a variety of the more common techniques have not been included in our book. The protocols described, based on rather sophisticated techniques for in vivo and in vitro studies, consist of molecular biology, biochemistry, and various types of immunoassays. Indeed, the authors have successfully accomplished an arduous task by presenting several topics in the simplest possible manner. We are confident that Cell Cycle Control and Dysregulation Protocols will facilitate and optimize the work of practical scientists involved in researching the cell cycle. We greatly acknowledge the extraordinary contribution of the authors in writing this book.

Recenzje

"State-of-the-art and highly practical...offers pathbreaking researchers powerful tools for probing the cell cycle regulation across a wide variety of pathologies." - Tumori

Spis treści

Part I. Analysis of Cyclins and Cyclin-Dependent Kinases The Biology of Cyclins and Cyclin-Dependent Protein Kinases: An Introduction Lucio Miele In Situ Immunofluorescence Analysis: Immunofluorescence Microscopy Amjad Javed, Sayyed K. Zaidi, Soraya E. Gutierrez, Christopher J. Lengner, Kimberly S. Harrington, Hayk Hovhannisyan, Brian C. Cho, Jitesh Pratap, Shirwin M. Pockwinse, Martin Montecino, André J. van Wijnen, Jane B. Lian, Janet L. Stein, and Gary S. Stein In Situ Immunofluorescence Analysis: Analyzing RNA Synthesis by 5-Bromouridine-5'-Triphosphate Labeling Amjad Javed, Sayyed K. Zaidi, Soraya E. Gutierrez, Christopher J. Lengner, Kimberly S. Harrington, Hayk Hovhannisyan, Brian C. Cho, Jitesh Pratap, Shirwin M. Pockwinse, Martin Montecino, André J. van Wijnen, Jane B. Lian, Janet L. Stein, and Gary S. Stein Immunofluorescence Analysis Using Epitope-Tagged Proteins: In Vitro System Amjad Javed, Sayyed K. Zaidi, Soraya E. Gutierrez, Christopher J. Lengner, Kimberly S. Harrington, Hayk Hovhannisyan, Brian C. Cho, Jitesh Pratap, Shirwin M. Pockwinse, Martin Montecino, André J. van Wijnen, Jane B. Lian, Janet L. Stein, and Gary S. Stein Analysis of In Vivo Gene Expression Using Epitope-Tagged Proteins Amjad Javed, Sayyed K. Zaidi, Soraya E. Gutierrez, Christopher J. Lengner, Kimberly S. Harrington, Hayk Hovhannisyan, Brian C. Cho, Jitesh Pratap, Shirwin M. Pockwinse, Martin Montecino, André J. van Wijnen, Jane B. Lian, Janet L. Stein, and Gary S. Stein Chromatin Immunoprecipitation Amjad Javed, Sayyed K. Zaidi, Soraya E. Gutierrez, Christopher J. Lengner, Kimberly S. Harrington, Hayk Hovhannisyan, Brian C. Cho, Jitesh Pratap, Shirwin M. Pockwinse, Martin Montecino, André J. van Wijnen, Jane B. Lian, Janet L. Stein, and Gary S. Stein Protein-Deoxyribonucleic Acid Interactions Linked to Gene Expression: Electrophoretic Mobility Shift Assay AmjadJaved, Sayyed K. Zaidi, Soraya E. Gutierrez, Christopher J. Lengner, Kimberly S. Harrington, Hayk Hovhannisyan, Brian C. Cho, Jitesh Pratap, Shirwin M. Pockwinse, Martin Montecino, André J. van Wijnen, Jane B. Lian, Janet L. Stein, and Gary S. Stein Protein-Deoxyribonucleic Acid Interactions Linked to Gene Expression: DNase I Digestion Amjad Javed, Sayyed K. Zaidi, Soraya E. Gutierrez, Christopher J. Lengner, Kimberly S. Harrington, Hayk Hovhannisyan, Brian C. Cho, Jitesh Pratap, Shirwin M. Pockwinse, Martin Montecino, André J. van Wijnen, Jane B. Lian, Janet L. Stein, and Gary S. Stein Protein-Deoxyribonucleic Acid Interactions Linked to Gene Expression: Ligation-Mediated Polymerase Chain Reaction Amjad Javed, Sayyed K. Zaidi, Soraya E. Gutierrez, Christopher J. Lengner, Kimberly S. Harrington, Hayk Hovhannisyan, Brian C. Cho, Jitesh Pratap, Shirwin M. Pockwinse, Martin Montecino, André J. van Wijnen, Jane B. Lian, Janet L. Stein, and Gary S. Stein Assays for Cyclin-Dependent Kinase Inhibitors Adrian M. Senderowicz. Protein Degradation Via the Proteosome, Henry Hoff, Hong Zhang, and Christian Sell Part II. Analysis of the Factors Involved in Cell Cycle Deregulation The Transformed Phenotype Henry Hoff, Barbara Belletti, Hong Zhang, and Christian Sell A Morphologic Approach to Detect Apoptosis Based on Electron Microscopy Martyn K. White and Caterina Cinti Detection of Apoptotic Deoxyribonucleic Acid Break by In Situ Nick Translation Carmela Trimarchi, Dario La Sala, Alessandra Zamparelli, and Caterina Cinti Part III. Cellular Response to Deoxyribonucleic Acid Damage Induction of Deoxyribonucleic Acid Damage by Alkylating Agents Salvatore Cortellino, David P. Turner, Domenico Albino, and Alfonso Bellacosa Induction of Deoxyribonucleic Acid Damage by g Irradiation Salvatore Cortellino, David P. Turner and Alfonso

Blurb

The dysregulation of the normal cell cycle underlies an enormous number of pathological conditions, including the neurodegenerative disorders, all cancers, and diabetes. In Cell Cycle Control and Dysregulation Protocols: Cyclins, Cyclin-Dependent Kinases, and Other Factors, expert laboratorians detail emerging methodologies for studying the cell cycle, its kinases, and kinase inhibitors. The authors focus on issues of gene expression in vivo and in vitro, the analysis of cyclin-dependent kinase inhibitors, protein degradation mediated by the proteosome, the analysis of the transformed cell phenotype, and innovative techniques to detect apoptosis. Many of the protocols are new, such as electron microsocopy to detect apoptosis and proteosome-mediated degradation, whereas others study the interactions between protein-protein, protein-DNA, and protein-RNA. The protocols follow the successful Methods in Molecular Biology™ series format, each one 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, Cell Cycle Control and Dysregulation Protocols: Cyclins, Cyclin-Dependent Kinases, and Other Factors offers pathbreaking researchers powerful tools for probing the cell cycle regulation across a wide variety of pathologies.