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This volume covers the mechanisms of pRb inactivation detailing repressive mechanisms commonly associated to cancer, and representative of the experimentally relevant tests used in the establishment of cancer diagnosis and prognosis. Chapters contain protocols and in-depth discussions for commonly used experimental approaches to assess the status and function of components of the pRb pathway, including pRb itself, in cell lines and biological samples.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.
Authoritative and practical, The Retinoblastoma Protein aims to serve as a guide to assist molecular cancer biologists in their search for understanding of the molecular functions of this preeminent tumorsuppressor.
List of contents
Characterization of RB1 Deletions in Interphase and Metaphase by Molecular Cytogenetics Exemplified in Chronic Lymphatic Leukemia.- Detection of RB1 Gene Copy Number Variations using a Multiplex Ligation-Dependent Probe Amplification Method.- A Fluorescent Quantitative Multiplex PCR Method to Detect Copy Number Changes in the RB1 Gene.- Using Methylation-specific PCR to study RB1 Promoter Hypermethylation.- Detection of Aberrant DNA Methylation Patterns in the RB1 Gene.- Detection of Retinoblastoma Protein Phosphorylation by Immunoblot Analysis.- Immunohistochemical Detection of the Retinoblastoma Protein.- Immunohistochemical Detection of Retinoblastoma Protein Phosphorylation in Human Tumor Samples.- Detection of CCND1 Locus Amplification by Fluorescence in situ Hybridization.- Detection of CCND1 Gene Copy Number Variations using Multiplex Ligation-dependent Probe Amplification and Fluorescence in situ Hybridization Methods.- Detection of p16 Promoter Hypermethylation by Methylation-specific PCR.- Immunohistochemical Detection of p16 in Clinical Samples.- Detection of E2F-DNA Complexes Using Chromatin Immunoprecipitation Assays.- Detection of E2F-induced Transcriptional Activity Using a Dual Luciferase Reporter Assay.- Detection of HPV E6/E7 mRNA in Clinical Samples using RNA in situ Hybridization.- CRISPR/Cas9 Mediated Knockout of Rb1 in Xenopus tropicalis.
Summary
This volume covers the mechanisms of pRb inactivation detailing repressive mechanisms commonly associated to cancer, and representative of the experimentally relevant tests used in the establishment of cancer diagnosis and prognosis. Chapters contain protocols and in-depth discussions for commonly used experimental approaches to assess the status and function of components of the pRb pathway, including pRb itself, in cell lines and biological samples.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.
Authoritative and practical, The Retinoblastoma Protein aims to serve as a guide to assist molecular cancer biologists in their search for understanding of the molecular functions of this preeminent tumorsuppressor.
