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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 tumor suppressor.

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1. Characterization of RB1 Deletions in Interphase and Metaphase by Molecular Cytogenetics Exemplified in Chronic Lymphatic Leukemia

Thomas Liehr

2. Detection of RB1 Gene Copy Number Variations using a Multiplex Ligation-Dependent Probe Amplification Method

Nejat Dalay

3. A Fluorescent Quantitative Multiplex PCR Method to Detect Copy Number Changes in the RB1 Gene

Chitra Kannabiran

4. Using Methylation-specific PCR to study RB1 Promoter Hypermethylation

Thaís M. McCormick and Maria da Glória da C. Carvalho
5. Detection of Aberrant DNA Methylation Patterns in the RB1 Gene

Sumadi Lukman Anwar and Ulrich Lehmann

6. Detection of Retinoblastoma Protein Phosphorylation by Immunoblot Analysis
Pedro G. Santiago-Cardona, Jaileene Pérez-Morales, and Jonathan González-Flores
7. Immunohistochemical Detection of the Retinoblastoma Protein

Charles A. Ishak, Matthew J. Cecchini, Christopher J. Howlett, and Frederick A. Dick

8. Immunohistochemical Detection of Retinoblastoma Protein Phosphorylation in Human Tumor Samples

Jaileene Pérez-Morales, Angel Nú¤ez-Marrero, and Pedro G. Santiago-Cardona
9. Detection of CCND1 Locus Amplification by Fluorescence in situ Hybridization

Margit Balázs, Viktória Koroknai, István Szász, and Szilvia Ecsedi

10. Detection of CCND1 Gene Copy Number Variations using Multiplex Ligation-dependent Probe Amplification and Fluorescence in situ Hybridization Methods

Akishi Ooi and Takeru Oyama

11. Detection of p16 Promoter Hypermethylation by Methylation-specific PCR

Javed Hussain Choudhury, Raima Das, Shaheen Laskar, Sharbadeb Kundu, Manish Kumar,

Partha Pratim Das, Yashmin Choudhury, Rosy Mondal, and Sankar Kumar Ghosh

12. Immunohistochemical Detection of p16 in Clinical Samples

Georgia Karpathiou and Michel Peoc´h

13. Detection of E2F-DNA Complexes Using Chromatin Immunoprecipitation Assays

Miyoung Lee, Lorraine J. Gudas, and Harold I. Saavedra

14. Detection of E2F-induced Transcriptional Activity Using a Dual Luciferase Reporter Assay

Ainhoa Iglesias-Ara, Nerea Osinalde, and Ana M. Zubiaga
15. Detection of HPV E6/E7 mRNA in Clinical Samples using RNA in situ Hybridization

Manishkumar Pandey, Priyanka G. Bhosale, and Manoj B. Mahimkar

16. CRISPR/Cas9 Mediated Knockout of Rb1 in Xenopus tropicalis

Thomas Naert and Kris Vleminckx

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