Cell Cycle Checkpoints, 1st ed. 2021 Methods and Protocols Methods in Molecular Biology Series, Vol. 2267

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.

Preface…

Table of Contents…

Contributing Authors..

1. Determination of CHK1 Cellular Localization by Immunofluorescence Microscopy

Yu-Che Cheng and Sheau-Yann Shieh

2. Detection of Post-Translationally Modified p53 by Western Blotting

Anna Estevan Barber and David W. Meek

3. Global Analyses to Identify Direct Transcriptional Targets of p53

Matthew D. Galbraith, Zdenek Andrysik, Kelly D. Sullivan, and Joaquín M. Espinosa

4. Analysis of Replication Dynamics Using the Single-Molecule DNA Fiber Spreading Assay

Stephanie Biber and Lisa Wiesmüller

5. Measuring Translation Efficiency by RNA Immunoprecipitation of Translation Initiation Factors

Chris Lucchesi, Shakur Mohibi, and Xinbin Chen

6. DNA Affinity Purification: A Pulldown Assay for Identifying and Analyzing Proteins Binding to Nucleic Acids

7. A Novel Strategy to Track Lysine-48 Ubiquitination by Fluorescence Resonance Energy Transfer

Kenneth Wu and Zhen-Qiang Pan

8. The DNA Damage Response in Xenopus laevis Cell-Free Extracts

Tomas Aparicio Casado and Jean Gautier

9. Mammalian Cell Fusion Assays for the Study of Cell Cycle Progression by Functional Complementation

Jongkuen Lee and David Dominguez-Siola

10. Knockdown of Target Genes by siRNA In Vitro

Songhee Back and James J. Manfredi

Ramy Rahme

12. Using TUNEL Assay to Quantitate p53-Induced Apoptosis in Mouse Tissues

Lois Resnick-Silverman

13. Generation and Analysis of dsDNA Breaks for Checkpoint and Repair Studies in Fission Yeast

Rohana Ramalingam and Matthew J. O’Connell

14. Calreticulin Exposure in Mitotic Catastrophe

Lucillia Bezu, Oliver Kepp, and Guido Kroemer

15. Quantification of eIF2α Phosphorylation Associated with Mitotic Catastrophe by Immunofluorescence Microscopy

Juliette Humeau, Lucillia Bezu, Oliver Kepp, Laura Senovilla, Peng Liu, and Guido Kroemer

16. Clonogenic Assays to Detect Cell Fate in Mitotic Catastrophe

José Manuel Bravo-San Pedro, Oliver Kepp, Allan Sauvat, Santiago Rello-Varona, Guido Kroemer, and Laura Senovilla

Subject Index List…