Part I. Preparation of Cell and Tissue Samplesn n Breast Tissue Banking: Collection, Handling, Storage, and Release of Tissue for Breast Cancer Researchn Linda Snell and Peter H. Watsonn n Frozen Biopsy Collection and Storage: Frozen Biopsy Samplesn Russell Leekn n Serial Biopsies/Fine-Needle Aspirates and Their Assessmentn Irene Boeddinghaus and Stephen R. D. Johnsonn n Breast Tissue Microarraysn Robert J. Springall and Cheryl E. Gillettn n Preparation of Tumor Homogenates for Subsequent Preparation of Cytosols, Membrane Fractions, RNA, and DNAn Naomi Robertson and Russell Leekn n Isolation of RNA From Tumor Samples: Single-Step Guanidinium Acid-Phenol Methodn Naomi Robertson and Russell Leekn n Isolation of DNA From Tumor Samplesn Naomi Robertson and Russell Leekn n Laser-Assisted Microdissection and Isolation of DNA and RNAn Ulrich Lehmann and Hans Kreipen n Part II. Markers of Clinical Outcome and Prognosisn n Traditional and Established Indicators of Prognosis and Treatment Successn Derek E. Roskell and Ian D. Buleyn n Sentinel Lymph Node Biopsy in Breast Cancern Celia Chao and Kelly M. McMastersn n Pathological Evaluation of Axillary Sentinel Lymph Nodes in Breast Cancern Michael Z. Gilcrease and Aysegul Sahinn n Measurement of Estrogen Receptor Status by Immunocytochemistry in Paraffin Wax Sectionsn Bharat Jasani, Anthony Douglas-Jones, Anthony Rhodes, Susan Wozniak, Peter J. Barrett-Lee, Julia Gee, and Robert Nicholsonn n Markers of Apoptosisn Fazlul H. Sarkar and Yiwei Lin n Quantitative Angiogenesis in Breast Cancern Stephen B. Foxn n Part III. Analysis of Tumor-Derived Proteins and Antigensn n Immunohistochemistryn Cheryl E. Gillettn n Detection of Aberrant Glycosylation in Breast Cancer Using Lectin Histochemistryn Tracey M. Carter and Susan A. Brooksn n SDS-PAGE and Western Blotting to Detect Proteins and Glycoproteins of Interest in Breast Cancer Researchn Chloe Osborne and Susan A. Brooksn n Breast Cancer Proteomics Using Two-Dimensional Electrophoresis: Studying the Breast Cancer Proteomen Miriam V. Dwek and Sarah L. Rawlingsn n Procedures for the Quantitative Protein Determination of Urokinase and Its Inhibitor, PAI-1, in Human Breast Cancer Tissue Extracts by ELISAn Manfred Schmitt, Alexandra S. Sturmheit, Anita Welk, Christel Schnelldorfer, and Nadia Harbeckn n Part IV. Analysis of Genes and Gene Expression in Tumor Specimensn n Fluorescence In Situ Hybridization and Comparative Genomic Hybridizationn Patricia Gorman and Rebecca Roylancen n Fluorescence In Situ Hybridization Assessment of c-myc Gene Amplification in Breast Tumor Tissuesn Jan K. Blancato, Mary Steele Williams, and Robert B. Dicksonn n Detection of HER2 Gene Amplification by Fluorescence In Situ Hybridization in Breast Cancern John M. S. Bartlett and Amanda Forsythn n In Situ Hybridization Combined With Immunohistochemistry to Localize Gene Expressionn Rosemary Jeffery, Toby Hunt, and Richard Poulsomn n Quantitation of RNA by Ribonuclease Protection Assayn John W. Mooren n Identification of Steroid Hormone-Regulated Genes in Breast Cancern Bruce R. Westley and Felicity E. B. Mayn n Sequencing of the Tumor Suppressor Gene TP 53n Barbro Linderholm, Torbjörn Norberg, and Jonas Berghn n Expression Profiling Using cDNA Microarraysn Chris Jones, Peter Simpson, Alan Mackay, and Sunil R. Lakhanin n Gene Expression Analysis Using Filter cDNA Microarraysn Peter Simpson, Chris Jones, Alan Mackay, and Sunil R. Lakhanin n Part V. Studying Cancer Cell Behavior In Vitro and In Vivon n Methods to Analyze the Effects of the Urokinase System on Cancer Cell Adhesion, Proliferation, Migration,
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.