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Full Description
This volume details widely used and newer lab protocols for studying hypoxic responses in physiology and diseases. Chapters guide the reader through the application of hypoxic conditions, to the techniques of molecular biology, biochemical, cell biology, genomic,bioinformatic, metabolic, and animal studies. 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 cutting-edge, Hypoxia: Methods and Protocols aims to provide a valuable set of tools that can be used to study hypoxia and beyond.
Contents
Whole Genome Sequencing-based Discovery of Structural Variants in Glioblastoma.- Whole Genome DNA Methylation Analysis of Human Glioblastoma using Illumina Bead Arrays.- Establishing Primary Human Glioblastoma Adherent Cultures from Operative Specimens.- Establishing Primary Human Glioblastoma Tumorsphere Cultures from Operative Specimens.- Isolation of GBM Stem Cells with Flow Cytometry.- Lentiviral Transduction of Primary Human Glioblastoma Cultures.- Selective Targeting of CD133-expressing Glioblastoma Stem Cells using Lentiviral Vectors.- Intracellular pH Measurements in Glioblastoma Cells using the pH-sensitive Dye BCECF.- Induction and Assessment of Hypoxia in Glioblastoma Cells in vitro.- Metabolomic Analysis of Glioma Cells Using Nanoflow Liquid Chromatography-tandem Mass Spectrometry.- Sample Preparation for Relative Quantitation of Proteins Using Tandem Mass Tags (TMT) and Mass Spectrometry (MS).- Single-Cell RNA Sequencing of Glioblastoma Cells.- Evaluation of Radioresponse and Radiosensitizers in Glioblastoma Organotypic Cultures.- Orthotopic Patient-derived Glioblastoma Xenografts in Mice.- Bioluminescent In Vivo Imaging of Orthotopic Glioblastoma Xenografts in Mice.- Evaluation of Vascularity, Blood Perfusion, and Oxygen Tension in Tumor Xenografts with Fluorescent Microscopy.- Probing Glioblastoma Tissue Heterogeneity with Laser Capture Microdissection.- Flow Cytometric Identification of Tumor-infiltrating Lymphocytes from Glioblastoma.- Modeling Glioma with Human Embryonic Stem Cell-derived Neural Lineages.
