Umfang:
Online-Ressource (XIX, 358 S.)
,
Ill., graph. Darst.
Ausgabe:
Online-Ausg.
ISBN:
9780128013298
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0128014830
,
9780128014837
Serie:
Methods in enzymology 543
Inhalt:
This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers research methods providing a a theoretical overview on metabolic alterations of cancer cells and a series of protocols that can be employed to study oncometabolism, in vitro, ex vivo and in vivo. Malignant cells exhibit metabolic changes when compared to their normal counterparts, owing to both genetic and epigenetic alterations. Although such a metabolic rewiring has recently been indicated as ""yet another"" general hallmark of canc
Anmerkung:
Description based upon print version of record
,
Cover image; Title page; Table of Contents; Copyright; Contributors; Preface; Acknowledgments; Chapter One: Methods to Measure Cytoplasmic and Mitochondrial Ca2 + Concentration Using Ca2 +-Sensitive Dyes; Abstract; 1 Introduction; 2 Experimental Components and Considerations; 3 Isolation and Differentiation of Naïve CD4+ CD25− T Cells from Spleen and Lymph Nodes of Wild-Type and Orai1−/− Mice; 4 Mitochondrial Ca2 + Measurement; 5 Data Handling and Processing; 6 Notes; 7 Conclusion; Acknowledgments
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Chapter Two: Methods to Measure Intracellular Ca2 + Fluxes with Organelle-Targeted Aequorin-Based ProbesAbstract; 1 Introduction; 2 Experimental Components and Considerations; 3 Application Examples; 4 Aequorin Improvement by Fusion with GFP; 5 Monitoring Ca2 + Fluxes in Living Organism by BRET; Acknowledgments; Chapter Three: Measuring Baseline Ca2 + Levels in Subcellular Compartments Using Genetically Engineered Fluorescent Indicators; Abstract; 1 Introduction; 2 Evolution of Recombinant Ca2 + Probes and Organelle Targeting; 3 Enhancing the Fluorophores
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4 Modulating Ca2 +-Binding Properties5 Developing Organelle Targeting; 6 Quantitative Considerations of Measurements of Organelle-Targeted Probes; 7 Experimental Protocols; 8 Semiquantitative Imaging of Basal Mitochondrial [Ca2 +] with a Novel Mitochondria-Targeted GCaMP6m (2mtGCaMP6m); 9 Semiquantitative Imaging of Agonist-Induced Mitochondrial Ca2 + Signals in Primary Neurons Using the Low-Affinity 2mtD4cpv Probe; 10 Semiquantitative Imaging of Basal, Steady-State [Ca2 +] in the ER; 11 Concluding Remarks; Chapter Four: Autophagy and Autophagic Flux in Tumor Cells; Abstract; 1 Introduction
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2 Autophagy Initiation and Autophagic Flux3 Autophagic Flux and Degradation; 4 Conclusion; Acknowledgments; Chapter Five: Methods to Assess Autophagy In Situ-Transmission Electron Microscopy Versus Immunohistochemistry; Abstract; 1 Introduction; 2 Transmission Electron Microscopy; 3 Immunohistochemistry; 4 Concluding Remarks; Acknowledgments; Chapter Six: Methods to Measure the Enzymatic Activity of PI3Ks; Abstract; 1 Introduction; 2 Analysis of PI3K Lipid Kinase Activity in Immunoprecipitate; 3 Analysis of the Activity of Small Molecule PI3K Inhibitors
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4 Analysis of the Localized Synthesis of PI3K Lipid Products5 Materials; Acknowledgments; Chapter Seven: Luciferase-Based Reporter to Monitor the Transcriptional Activity of the SIRT3 Promoter; Abstract; 1 Introduction; 2 Protocol for Construction of Reporter Plasmid; 3 Assaying SIRT3 Promoter Activity with the Construct; 4 Verifying Reporter Results with Complementary Methods; 5 Summary; Acknowledgments; Chapter Eight: Metabolomic Profiling of Cultured Cancer Cells; Abstract; 1 Introduction; 2 Cell Culture, Treatments, and Sample Collection
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3 Chromatography, Mass Spectrometry, and Data Analysis
Weitere Ausg.:
Erscheint auch als Druck-Ausgabe Cell-wide Metabolic Alterations Associated With Malignancy
Sprache:
Englisch
Fachgebiete:
Biologie
Schlagwort(e):
Carcinogenese
;
Primärstoffwechsel
;
Electronic books
;
Aufsatzsammlung
Mehr zum Autor:
Kroemer, Guido 1961-
Mehr zum Autor:
Galluzzi, Lorenzo 1980-
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