Cell Cycle Control and Dysregulation Protocols

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.

Includes supplementary material: sn.pub/extras

Klappentext
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.

Inhalt
Analysis of Cyclins and Cyclin-Dependent Kinases.- The Biology of Cyclins and Cyclin-Dependent Protein Kinases.- In Situ Immunofluorescence Analysis.- In Situ Immunofluorescence Analysis.- Immunofluorescence Analysis Using Epitope-Tagged Proteins.- Analysis of In Vivo Gene Expression Using Epitope-Tagged Proteins.- Chromatin Immunoprecipitation.- Protein-Deoxyribonucleic Acid Interactions Linked to Gene Expression.- Protein-Deoxyribonucleic Acid Interactions Linked to Gene Expression.- Protein-Deoxyribonucleic Acid Interactions Linked to Gene Expression.- Assays for Cyclin-Dependent Kinase Inhibitors.- Protein Degradation Via the Proteosome.- Analysis of the Factors Involved in Cell Cycle Deregulation.- The Transformed Phenotype.- A Morphologic Approach to Detect Apoptosis Based on Electron Microscopy.- Detection of Apoptotic Deoxyribonucleic Acid Break by In Situ Nick Translation.- Cellular Response to Deoxyribonucleic Acid Damage.- Induction of Deoxyribonucleic Acid Damage by Alkylating Agents.- Induction of Deoxyribonucleic Acid Damage by ? Irradiation.- Ultraviolet Irradiation of Cells.- Retroviridae-Based Vectors: Protocols for Lentiviral- and Retroviral-Mediated Gene Transfer to Engineer Cell Culture Systems.- Transient Production of Retroviral- and Lentiviral-Based Vectors for the Transduction of Mammalian Cells.- Determination of Functional Viral Titer by Drug-Resistance Colony Assay, Expression of Green Fluorescent Protein, and ?-Galactoside Staining.- Retroviral and Lentiviral Vector Titration by the Analysis of the Activity of Viral Reverse Transcriptase.- Detection of Gene Expression in Subcellular Compartments.- Single and Double Colloidal Gold Labeling in Postembedding Immunoelectron Microscopy.- Multifluorescence Labeling and Colocalization Analyses.