دانلود کتاب روش ها و پروتکل های زیست شناسی سلول سرطانی

This volume provides detailed methods for the dissection of mechanisms underlying cancer cell biology. This book begins with fundamental techniques and considerations for culturing cancer cell lines and then more advanced techniques to study patient-derived tissue using xenografts and cryopreservation of tumor cells. The book then describes methods to analyze the co-culture of breast cancer cells with adipocytes in two-dimensional and threedimensional systems. Next are a series of chapters that allows the researcher to analyze cancer cell physiology including cell proliferation, protein secretion by ELISA, flow cytometry-based detection of protein levels by conventional fluorescence and mass spectrometry, and multi-parametric immunofluorescence analysis of tumor tissue. Also included are analysis of protein expression by western blot, analysis of surface protein levels and recycling by biotinylation assay, and proteomics analysis by liquid chromatography mass spectrometry. This is followed by chapters describing a sophisticated analysis of cellular metabolism using the Seahorse assay, specific analysis of cancer cell mutations by CRISPR/ Cas9, single cell RNA sequencing, along with a chapter on validation by reverse transcriptase quantitative PCR, and then bioinformatics analysis of RNA-seq data. Lastly, chapters describing the isolation and characterization of extracellular vesicles follow. This volume follows the highly successful Methods in Molecular Biology format, with each chapter including an introduction to their respective techniques; lists of the necessary materials and reagents; step-by-step, readily reproducible laboratory protocols; and tips on troubleshooting and avoiding known pitfalls.