Main description:

This volume explores the latest advancements in the field of cell cycle checkpoints and their implications for human diseases. Chapters in this book cover topics such as post-translationally modified p53 by western blotting; CHK1 cellular localization by immunofluorescence microscopy; DNA affinity purification; knockdown of target genes by siRNA in vitro; and calreticulin exposure in mitotic catastrophe. 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 tips on troubleshooting and avoiding known pitfalls.

Cutting-edge and thorough, Cell Cycle Checkpoints: Methods and Protocols is a valuable resource for researcher interested in learning more about this developing field.

Contents:

Preface...Table of Contents...Contributing Authors..
1. Determination of CHK1 Cellular Localization by Immunofluorescence MicroscopyYu-Che Cheng and Sheau-Yann Shieh
2. Detection of Post-Translationally Modified p53 by Western BlottingAnna Estevan Barber and David W. Meek
3. Global Analyses to Identify Direct Transcriptional Targets of p53Matthew D. Galbraith, Zdenek Andrysik, Kelly D. Sullivan, and Joaquin M. Espinosa
4. Analysis of Replication Dynamics Using the Single-Molecule DNA Fiber Spreading AssayStephanie Biber and Lisa Wiesmuller
5. Measuring Translation Efficiency by RNA Immunoprecipitation of Translation Initiation FactorsChris Lucchesi, Shakur Mohibi, and Xinbin Chen
6. DNA Affinity Purification: A Pulldown Assay for Identifying and Analyzing Proteins Binding to Nucleic AcidsGerd A. Muller and Kurt Engeland
7. A Novel Strategy to Track Lysine-48 Ubiquitination by Fluorescence Resonance Energy TransferKenneth Wu and Zhen-Qiang Pan
8. The DNA Damage Response in Xenopus laevis Cell-Free ExtractsTomas Aparicio Casado and Jean Gautier
9. Mammalian Cell Fusion Assays for the Study of Cell Cycle Progression by Functional ComplementationJongkuen Lee and David Dominguez-Siola
10. Knockdown of Target Genes by siRNA In VitroSonghee Back and James J. Manfredi
11. Assaying Cell Cycle Status Using Flow CytometryRamy Rahme
12. Using TUNEL Assay to Quantitate p53-Induced Apoptosis in Mouse TissuesLois Resnick-Silverman
13. Generation and Analysis of dsDNA Breaks for Checkpoint and Repair Studies in Fission YeastRohana Ramalingam and Matthew J. O'Connell
14. Calreticulin Exposure in Mitotic CatastropheLucillia Bezu, Oliver Kepp, and Guido Kroemer
15. Quantification of eIF2 Phosphorylation Associated with Mitotic Catastrophe by Immunofluorescence MicroscopyJuliette Humeau, Lucillia Bezu, Oliver Kepp, Laura Senovilla, Peng Liu, and Guido Kroemer
16. Clonogenic Assays to Detect Cell Fate in Mitotic CatastropheJose Manuel Bravo-San Pedro, Oliver Kepp, Allan Sauvat, Santiago Rello-Varona, Guido Kroemer, and Laura Senovilla
Subject Index List...