This volume focuces on emerging methodologies for studying the cell cycle, kinases and kinases inhibitors.

Expert laboratorians detail emerging methodologies for studying the cell cycle, its kinases, and kinase inhibitors. The authors focus the 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.

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, Andre 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, Andre 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, Andre 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, Andre 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, Andre 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
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, Andre 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, Andre 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, Andre 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 Bellacosa
Ultraviolet Irradiation of Cells
David P. Turner, Anthony T. Yeung, and Alfonso Bellacosa
Part IV. 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
Tiziana Tonini, Pier Paolo Claudio, Antonio Giordano, and Gaetano Romano
Determination of Functional Viral Titer by Drug-Resistance Colony Assay, Expression of Green Fluorescent Protein, and b-Galactoside Staining
Tiziana Tonini, Pier Paolo Claudio, Antonio Giordano, and Gaetano Romano
Retroviral and Lentiviral Vector Titration by the Analysis of the Activity of Viral Reverse Transcriptase
Tiziana Tonini, Pier Paolo Claudio, Antonio Giordano, and Gaetano Romano
Part V. Detection of Gene Expression in Subcellular Compartments
Single and Double Colloidal Gold Labeling in Postembedding Immunoelectron Microscopy
Nicoletta Zini, Liliana Solimando, Caterina Cinti, and Nadir Mario Maraldi
Multifluorescence Labeling and Colocalization Analyses
Massimo Riccio, Maja Dembic, Caterina Cinti, and Spartaco Santi
Index