Breast Cancer Research Protocols von Susan A Brooks/Adrian Harris

A collection of both well-established and cutting-edge methods for investigating breast cancer biology not only in the laboratory, but also in clinical settings. These readily reproducible techniques solve a variety of problems, ranging from how to collect, store, and prepare human breast tumor samples for analysis, to analyzing cells in vivo and in vitro. Additional chapters address the technology of handling biopsies, new methods for analyzing genes and gene expression, markers of clinical outcome and progress, analysis of tumor-derived proteins and antigens, validating targets, and investigating the biology of newly discovered genes.

InhaltsangabePart I. Preparation of Cell and Tissue Samples Breast Tissue Banking: Collection, Handling, Storage, and Release of Tissue for Breast Cancer Research Linda Snell and Peter H. Watson Frozen Biopsy Collection and Storage: Frozen Biopsy Samples Russell Leek Serial Biopsies/Fine-Needle Aspirates and Their Assessment Irene Boeddinghaus and Stephen R. D. Johnson Breast Tissue Microarrays Robert J. Springall and Cheryl E. Gillett Preparation of Tumor Homogenates for Subsequent Preparation of Cytosols, Membrane Fractions, RNA, and DNA Naomi Robertson and Russell Leek Isolation of RNA From Tumor Samples: Single-Step Guanidinium Acid-Phenol Method Naomi Robertson and Russell Leek Isolation of DNA From Tumor Samples Naomi Robertson and Russell Leek LaserAssisted Microdissection and Isolation of DNA and RNA Ulrich Lehmann and Hans Kreipe Part II. Markers of Clinical Outcome and Prognosis Traditional and Established Indicators of Prognosis and Treatment Success Derek E. Roskell and Ian D. Buley Sentinel Lymph Node Biopsy in Breast Cancer Celia Chao and Kelly M. McMasters Pathological Evaluation of Axillary Sentinel Lymph Nodes in Breast Cancer Michael Z. Gilcrease and Aysegul Sahin Measurement of Estrogen Receptor Status by Immunocytochemistry in Paraffin Wax Sections Bharat Jasani, Anthony Douglas-Jones, Anthony Rhodes, Susan Wozniak, Peter J. Barrett-Lee, Julia Gee, and Robert Nicholson Markers of Apoptosis Fazlul H. Sarkar and Yiwei Li Quantitative Angiogenesis in Breast Cancer Stephen B. Fox Part III. Analysis of Tumor-Derived Proteins and Antigens Immunohistochemistry Cheryl E. Gillett Detection of Aberrant Glycosylation in Breast Cancer Using Lectin Histochemistry Tracey M. Carter and Susan A. Brooks SDSPAGE and Western Blotting to Detect Proteins and Glycoproteins of Interest in Breast Cancer Research Chloe Osborne and Susan A. Brooks Breast Cancer Proteomics Using Two-Dimensional Electrophoresis: Studying the Breast Cancer Proteome Miriam V. Dwek and Sarah L. Rawlings Procedures for the Quantitative Protein Determination of Urokinase and Its Inhibitor, PAI-1, in Human Breast Cancer Tissue Extracts by ELISA Manfred Schmitt, Alexandra S. Sturmheit, Anita Welk, Christel Schnelldorfer, and Nadia Harbeck Part IV. Analysis of Genes and Gene Expression in Tumor Specimens Fluorescence In Situ Hybridization and Comparative Genomic Hybridization Patricia Gorman and Rebecca Roylance Fluorescence In Situ Hybridization Assessment of c-myc Gene Amplification in Breast Tumor Tissues Jan K. Blancato, Mary Steele Williams, and Robert B. Dickson Detection of HER2 Gene Amplification by Fluorescence In Situ Hybridization in Breast Cancer John M. S. Bartlett and Amanda Forsyth In Situ Hybridization Combined With Immunohistochemistry to Localize Gene Expression Rosemary Jeffery, Toby Hunt, and Richard Poulsom Quantitation of RNA by Ribonuclease Protection Assay John W. Moore Identification of Steroid Hormone-Regulated Genes in Breast Cancer Bruce R. Westley and Felicity E. B. May Sequencing of the Tumor Suppressor Gene TP 53 Barbro Linderholm, Torbjörn Norberg, and Jonas Bergh Expression Profiling Using cDNA Microarrays Chris Jones, Peter Simpson, Alan Mackay, and Sunil R. Lakhani Gene Expression Analysis Using Filter cDNA Microarrays Peter Simpson, Chris Jones, Alan Mackay, and Sunil R. Lakhani Part V. Studying Cancer Cell Behavior In Vitro and In Vivo Methods to Analyze the Effects of the Urokinase System on Cancer Cell Adhesion, Proliferation, Migration,

Part I. Preparation of Cell and Tissue Samples Breast Tissue Banking: Collection, Handling, Storage, and Release of Tissue for Breast Cancer Research Linda Snell and Peter H. Watson Frozen Biopsy Collection and Storage: Frozen Biopsy Samples Russell Leek Serial Biopsies/Fine-Needle Aspirates and Their Assessment Irene Boeddinghaus and Stephen R. D. Johnson Breast Tissue Microarrays Robert J. Springall and Cheryl E. Gillett Preparation of Tumor Homogenates for Subsequent Preparation of Cytosols, Membrane Fractions, RNA, and DNA Naomi Robertson and Russell Leek Isolation of RNA From Tumor Samples: Single-Step Guanidinium Acid-Phenol Method Naomi Robertson and Russell Leek Isolation of DNA From Tumor Samples Naomi Robertson and Russell Leek Laser-Assisted Microdissection and Isolation of DNA and RNA Ulrich Lehmann and Hans Kreipe Part II. Markers of Clinical Outcome and Prognosis Traditional and Established Indicators of Prognosis and Treatment Success Derek E. Roskell and Ian D. Buley Sentinel Lymph Node Biopsy in Breast Cancer Celia Chao and Kelly M. McMasters Pathological Evaluation of Axillary Sentinel Lymph Nodes in Breast Cancer Michael Z. Gilcrease and Aysegul Sahin Measurement of Estrogen Receptor Status by Immunocytochemistry in Paraffin Wax Sections Bharat Jasani, Anthony Douglas-Jones, Anthony Rhodes, Susan Wozniak, Peter J. Barrett-Lee, Julia Gee, and Robert Nicholson Markers of Apoptosis Fazlul H. Sarkar and Yiwei Li Quantitative Angiogenesis in Breast Cancer Stephen B. Fox Part III. Analysis of Tumor-Derived Proteins and Antigens Immunohistochemistry Cheryl E. Gillett Detection of Aberrant Glycosylation in Breast Cancer Using Lectin Histochemistry Tracey M. Carter and Susan A. Brooks SDS-PAGE and Western Blotting to Detect Proteins and Glycoproteins of Interest in Breast Cancer Research Chloe Osborne and Susan A. Brooks Breast Cancer Proteomics Using Two-Dimensional Electrophoresis: Studying the Breast Cancer Proteome Miriam V. Dwek and Sarah L. Rawlings Procedures for the Quantitative Protein Determination of Urokinase and Its Inhibitor, PAI-1, in Human Breast Cancer Tissue Extracts by ELISA Manfred Schmitt, Alexandra S. Sturmheit, Anita Welk, Christel Schnelldorfer, and Nadia Harbeck Part IV. Analysis of Genes and Gene Expression in Tumor Specimens Fluorescence In Situ Hybridization and Comparative Genomic Hybridization Patricia Gorman and Rebecca Roylance Fluorescence In Situ Hybridization Assessment of c-myc Gene Amplification in Breast Tumor Tissues Jan K. Blancato, Mary Steele Williams, and Robert B. Dickson Detection of HER2 Gene Amplification by Fluorescence In Situ Hybridization in Breast Cancer John M. S. Bartlett and Amanda Forsyth In Situ Hybridization Combined With Immunohistochemistry to Localize Gene Expression Rosemary Jeffery, Toby Hunt, and Richard Poulsom Quantitation of RNA by Ribonuclease Protection Assay John W. Moore Identification of Steroid Hormone-Regulated Genes in Breast Cancer Bruce R. Westley and Felicity E. B. May Sequencing of the Tumor Suppressor Gene TP 53 Barbro Linderholm, Torbjörn Norberg, and Jonas Bergh Expression Profiling Using cDNA Microarrays Chris Jones, Peter Simpson, Alan Mackay, and Sunil R. Lakhani Gene Expression Analysis Using Filter cDNA Microarrays Peter Simpson, Chris Jones, Alan Mackay, and Sunil R. L ...