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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.
Sommario
Determination of CHK1 Cellular Localization by Immunofluorescence Microscopy.- Detection of Post-Translationally Modified p53 by Western Blotting.- Global Analyses to Identify Direct Transcriptional Targets of p53.- Analysis of Replication Dynamics Using the Single-Molecule DNA Fiber Spreading Assay.- Measuring Translation Efficiency by RNA Immunoprecipitation of Translation Initiation Factors.- DNA Affinity Purification: A Pulldown Assay for Identifying and Analyzing Proteins Binding to Nucleic Acids.- A Novel Strategy to Track Lysine-48 Ubiquitination by Fluorescence Resonance Energy Transfer.- The DNA Damage Response in Xenopus laevis Cell-Free Extracts.- Mammalian Cell Fusion Assays for the Study of Cell Cycle Progression by Functional Complementation.- Knockdown of Target Genes by siRNA In Vitro.- Assaying Cell Cycle Status Using Flow Cytometry.- Using TUNEL Assay to Quantitate p53-Induced Apoptosis in Mouse Tissues.- Generation and Analysis of dsDNA Breaks for Checkpoint and Repair Studies in Fission Yeast.- Calreticulin Exposure in Mitotic Catastrophe.- Quantification of eIF2 Phosphorylation Associated with Mitotic Catastrophe by Immunofluorescence Microscopy.- Clonogenic Assays to Detect Cell Fate in Mitotic Catastrophe.
