Case Studies in Cell Biology

 
 
Academic Press
  • 1. Auflage
  • |
  • erschienen am 15. März 2016
  • |
  • 406 Seiten
 
E-Book | ePUB mit Adobe DRM | Systemvoraussetzungen
E-Book | PDF mit Adobe DRM | Systemvoraussetzungen
978-0-12-801684-8 (ISBN)
 

Case Studies in Cell Biology presents real world scenarios to help readers use science process and reasoning skills. The case studies require application and analyzation of concepts beyond rote memory of biological concepts.

The book is based on the student learning outcomes from the American Society for Cell Biology, offering practical application for both the classroom and research laboratory.


  • Guides the reader in applying knowledge directly to real world scenarios
  • Includes case studies to bridge foundational cell biological concepts to translational science
  • Aids students in synthesizing information and applying science processes
  • Englisch
  • San Diego
  • |
  • USA
Elsevier Science
  • 27,00 MB
978-0-12-801684-8 (9780128016848)
0128016841 (0128016841)
weitere Ausgaben werden ermittelt
  • Cover
  • Title Page
  • Copyright Page
  • Dedication
  • Contents
  • Biography
  • Preface
  • Chapter 1 - Introduction to Scientific Method from a Cellular Perspective
  • Subchapter 1.1 - Scientific Method in Action [1]
  • Introduction
  • Background
  • Methods
  • Cell cultures
  • Microinjection
  • Electron microscopy
  • Calculation of nuclear uptake and size distribution
  • Results
  • Reference
  • Chapter 2 - Cellular Biodiversity
  • Subchapter 2.1 - Cellular Biodiversity on the High Seas [1]
  • Introduction
  • Background
  • Methods
  • Cell culture and population density
  • Characterization of photosynthetic pigments
  • Results
  • Subchapter 2.2 - The Mystery of the Missing Mitochondria [2]
  • Introduction
  • Background
  • Methods
  • Subcellular fractionation of giardia
  • Transmission electron immunomicroscopy
  • Results
  • References
  • Chapter 3 - Proteins
  • Subchapter 3.1 - Bite of the Brown Recluse Spider: An Introduction to Protein Gel Electrophoresis [1]
  • Introduction
  • Background
  • Methods
  • Red blood cell ghosts
  • Venom digestion assays
  • Protein gel electrophoresis
  • Results
  • Subchapter 3.2 - Where to Start? A Case of Too Many AUGs [3]
  • Introduction
  • Background
  • Methods
  • Yeast culture conditions
  • CDC9-GFP fusions, CDC9 mutagenesis, and HA tagging
  • Subcellular fractionation and protease K digestion
  • Immunoblotting
  • Results
  • Subchapter 3.3 - Cotranslational Translocation: Gatekeepers of the RER [4]
  • Introduction
  • Background
  • Methods
  • In vitro translation
  • Crosslinking assay
  • Flotation assay
  • Protease resistance assay
  • Results
  • Subchapter 3.4 - From Sequence to Function [5]
  • Introduction
  • Background
  • Methods
  • Isolation and structural analysis of bovine AQP1
  • Data processing
  • Results
  • Subchapter 3.5 - Visualizing Protein Conformation [6]
  • Introduction
  • Background
  • Methods
  • Protein expression and purification
  • Electron microscopy
  • SNARE complex labeling
  • Results
  • Subchapter 3.6 - Second Chance Chaperones: How Misfolded Proteins get Refolded [7]
  • Introduction
  • Background
  • Methods
  • Purification of Hsp104
  • Purification of Hsp40 and Hsp70
  • Preparation of yeast lysates
  • Turbidity assay
  • Refolding assays
  • Gel filtration
  • Results
  • References
  • Chapter 4 - The Nucleus
  • Subchapter 4.1 - Nuclear Pore Complex Assembly is a House of Cards [1]
  • Introduction
  • Background
  • Methods
  • Nuclear reconstitution assay
  • Immunodepletion
  • Nuclear import
  • Immunoblotting
  • DNA replication assay
  • Scanning electron microscopy
  • Results
  • Subchapter 4.2 - Importin b: The Important Importin [2]
  • Introduction
  • Background
  • Methods
  • Preparation of a high-speed supernatant from Xenopus eggs
  • Immunodepletion of importin from cytosol
  • Nuclear protein import assay
  • Nuclear localization signal binding assay
  • Protein gel electrophoresis and immunoblotting
  • Results
  • Subchapter 4.3 - A Tale of tRNA Transport [3]
  • Introduction
  • Background
  • Methods
  • In vitro translation and 35S-labeling
  • Injection into Xenopus frog eggs
  • In vitro binding assays
  • Gel electrophoresis and immunoblotting
  • Results
  • Subchapter 4.4 - When It Comes to the Nucleus - Size Matters [4]
  • Introduction
  • Background
  • Methods
  • Xenopus egg extracts and nuclear assembly
  • Immunodepletions and recombinant proteins
  • Fluorescence microscopy and image analysis
  • Immunoblots
  • Results
  • References
  • Chapter 5 - Membranes and Membrane Transport
  • Subchapter 5.1 - Flip This Lipid [1]
  • Introduction
  • Background
  • Methods
  • Microsomes and submicrosomal membrane fractions
  • Preparation of salt-washed rough microsomal membranes
  • Reconstitution of proteoliposomes
  • Protease treatment of proteoliposomes
  • Transport of [3H]diC4PC into vesicles
  • Results
  • Subchapter 5.2 - Navigating the Bilayer: Lipid Rafts and Caveolae [2]
  • Introduction
  • Background
  • Methods
  • Plasma membrane purification
  • Sucrose density gradient centrifugation
  • Extraction of membrane lipid domains
  • Lipid analysis
  • Electron microscopy
  • Results
  • Subchapter 5.3 - Shifting Gears: Calcium Transport in Flagella [3]
  • Introduction
  • Background
  • Methods
  • Cell cultures
  • Reverse transcriptase - PCR of regenerating flagella
  • RNA interference suppression of CAV2 expression
  • CAV2 antibodies
  • Results
  • References
  • Chapter 6 - Cytoskeleton and Intracellular Motility
  • Subchapter 6.1 - Plakins: Keeping the Cytoskeleton Safe [1]
  • Introduction
  • Background
  • Methods
  • Mice
  • Transfections and immunofluorescence
  • Primary culturing of mouse neurons
  • In vitro microtubule-binding assays
  • Results
  • Subchapter 6.2 - The Moving Story of a Microtubule Motor Protein [2]
  • Introduction
  • Background
  • Methods
  • Preparation of dissociated axoplasm
  • Preparation of vesicles and supernatant from axoplasm
  • Preparation of squid optic lobe microtubules
  • In vitro motility assay
  • Electron microscopy
  • Results
  • Subchapter 6.3 - The WASP and the Barbed End [3]
  • Introduction
  • Background
  • Methods
  • Protein expression, purification, and fluorescent labeling
  • Attachment zone localization assay
  • Reconstitution of in vitro membrane motility
  • Actin filament capture assay
  • Results
  • Subchapter 6.4 - Cilia Grow Where Vesicles Go [4]
  • Introduction
  • Background
  • Methods
  • Cell culturing and extraction of cytoplasm
  • Isolation of cytoplasmic vesicles
  • Immunoelectron microscopy
  • Immunofluorescence microscopy
  • Results
  • References
  • Chapter 7 - Organelles
  • Subchapter 7.1 - Putting the "Retic" in the Endoplasmic Reticulum [1]
  • Introduction
  • Background
  • Methods
  • Vesicle isolation
  • In vitro network formation
  • Aequorin Ca2+ assay
  • Sulfhydryl modification and protein isolation
  • Results
  • Subchapter 7.2 - How the Golgi Stacks Up [2]
  • Introduction
  • Background
  • Methods
  • Membrane and cytosol isolation
  • Cell-free disassembly/reassembly assays
  • Protease and extraction treatments
  • Immunoblots and immunoprecipitation
  • Electron microscopy
  • Results
  • Subchapter 7.3 - Case of the Coated Vesicle [3]
  • Introduction
  • Background
  • Methods
  • Cell-free vesicle-budding assay
  • Cell-free vesicle fusion to Golgi membranes
  • Gel filtration chromatography
  • Electron microscopy and immunocytochemistry
  • Results
  • Subchapter 7.4 - How to Build a Peroxisome [4]
  • Introduction
  • Background
  • Methods
  • Yeast strains and DNA
  • Galactose induction for fluorescence pulse chase
  • Mating assays
  • Microscopy
  • Results
  • Subchapter 7.5 - Putting the Squeeze on Mitochondria [5]
  • Introduction
  • Background
  • Methods
  • Cell lines and labeling
  • Inhibition of Myosin II
  • Microscopy
  • Results
  • References
  • Chapter 8 - Exocytosis
  • Subchapter 8.1 - Coat Proteins and Vesicle Transport [1]
  • Introduction
  • Background
  • Methods
  • Cell culture and gene expression
  • Antibodies and immunofluorescence
  • Results
  • Subchapter 8.2 - Endomembrane Transport in the Absence of a Cell [2]
  • Introduction
  • Background
  • Methods
  • CHO cells and VSV
  • Homogenization and gradient purification of subcellular fractions
  • Cell-free system for Golgi transport
  • In vivo inhibition of protein synthesis
  • Results
  • Subchapter 8.3 - Extra Large Export: A Case for Cisternal Maturation [3]
  • Introduction
  • Background
  • Methods
  • Primary culture of chick embryo fibroblast cells
  • Immunofluorescence
  • Immunoelectron microscopy
  • PC "wave" protocols
  • Assay of PC release
  • Results
  • References
  • Chapter 9 - Endocytosis
  • Subchapter 9.1 - Following the Fate of a Phagosome [1]
  • Introduction
  • Background
  • Methods
  • InlA-coated beads
  • Determination of Alexa488-InlA bead internalization
  • Determination of acidification using FITC-InlA beads
  • Determination of phagosome/lysosome fusion
  • Kinetics of phagocytosis
  • Results
  • Subchapter 9.2 - Catching a Receptor by the Tail [2]
  • Introduction
  • Background
  • Methods
  • Cell culture
  • Immunoprecipitation
  • Surface plasmon resonance detection
  • Results
  • Subchapter 9.3 - Can Clathrin Bend a Membrane? [3]
  • Introduction
  • Background
  • Methods
  • Cell-free system for clathrin binding
  • Sedimentation assay
  • Electron microscopy
  • Results
  • Subchapter 9.4 - Modeling Membrane Fission [4]
  • Introduction
  • Background
  • Methods
  • SUPER templates
  • Dynamin protein
  • Dynamin assays
  • Results
  • References
  • Chapter 10 - Cell Walls and Cell Adhesion
  • Subchapter 10.1 - Biofilms and Antibiotic Resistance [1]
  • Introduction
  • Background
  • Methods
  • Bacterial strains
  • Antibiotic resistance
  • Results
  • Subchapter 10.2 - DIY ECM: Cortactin and the Secretion of Fibronectin [2]
  • Introduction
  • Background
  • Methods
  • Cells and cell culture
  • Single-cell motility assay
  • Lamellipodia persistence assay
  • Immunodectection procedures
  • Results
  • Subchapter 10.3 - Bundling the Brush Border [3]
  • Introduction
  • Background
  • Methods
  • Cell culture
  • Microscopy
  • Analysis of microvillar clustering
  • Bead aggregation assay
  • Results
  • References
  • Chapter 11 - Cell Metabolism
  • Subchapter 11.1 - When Glucose is Low, Something Must Go [1]
  • Introduction
  • Background
  • Methods
  • Yeast growth and carbon depletion
  • Measurement of ATP levels
  • Fluorescence recovery after photobleaching
  • Yeast cell permeabilization
  • Results
  • Subchapter 11.2 - Do Plants Really Need Two Photosystems? [2]
  • Introduction
  • Background
  • Methods
  • Algal strains and growth conditions
  • Gas exchange measurements
  • Immunoblotting
  • Results
  • Subchapter 11.3 - FREX: Opening a Window Into Cellular Metabolism [3]
  • Introduction
  • Background
  • Methods
  • Charactertization of Frex in vitro
  • Cell culture
  • Fluorescence microscopy
  • Live cell fluorescence measurements
  • Results
  • References
  • Chapter 12 - Cell Signaling
  • Subchapter 12.1 - How Cells Know When It's Time to Go [1]
  • Introduction
  • Background
  • Methods
  • Cell culture
  • Assays
  • Northern and immunoblot analysis
  • Results
  • Subchapter 12.2 - Can You "Ad" Hear Me Now? Signaling and Intraflagellar Transport [2]
  • Introduction
  • Background
  • Methods
  • Cells and cell culture
  • Fractionation of cells and flagella
  • Immunoblotting and immunofluorescence
  • In vitro protein kinase assay
  • Results
  • References
  • Chapter 13 - Cell Cycle
  • Subchapter 13.1 - Now You See It, Now You Don't: The Discovery of Cyclin [1]
  • Introduction
  • Background
  • Methods
  • Preparation and incubation of eggs and embryos
  • Sample preparation and analysis
  • Determination of the cleavage index
  • Results
  • Subchapter 13.2 - Sorting Out Cyclins [2]
  • Introduction
  • Background
  • Methods
  • Synchronization of HeLa cells
  • Tracking nuclear envelope breakdown using a fluorescent mitosis biosensor
  • Knockdown of cyclins by siRNA
  • Determination of ploidy by flow cytometry
  • Immunoblot analysis
  • Results
  • Subchapter 13.3 - Of Centriole Separation and Cyclins [3]
  • Introduction
  • Background
  • Methods
  • Frog embryo injections and fluorescent labeling
  • In vitro centriole separation assay
  • Embryo extract modifications
  • Results
  • Subchapter 13.4 - The Path to S Phase is Paved With Phosphorylation [4]
  • Introduction
  • Background
  • Methods
  • Plasmid constructs and transfection
  • CAT assay
  • Immunoprecipitation and Western blot
  • Results
  • References
  • Chapter 14 - Cell Division
  • Subchapter 14.1 - Push and Pull: How Motor Proteins Help Build a Spindle [1]
  • Introduction
  • Background
  • Methods
  • Cell culture and nocodazole treatment
  • Inhibitors
  • Time lapse microscopy
  • Results
  • Subchapter 14.2 - Ready, Set, Anaphase! [2]
  • Introduction
  • Background
  • Methods
  • Xenopus egg extracts
  • Histone H1 phosphorylation assay
  • Securin degradation
  • Immunofluorescence
  • Results
  • Subchapter 14.3 - Building Cell Walls - Cytokinesis in a Plant Cell [3]
  • Introduction
  • Background
  • Methods
  • Plant and cell material
  • Immunoprecipitation
  • Kinase assay
  • Immunofluorescence and light microscopy
  • Results
  • References
  • Chapter 15 - Cell Systems
  • Subchapter 15.1 - Do Bumblebees Have B cells? A Case of Insect Immunity [1]
  • Introduction
  • Background
  • Methods
  • Insects
  • Bacteria
  • Exposure combinations and survival
  • Exposure combinations and bacterial clearance
  • Results
  • Subchapter 15.2 - What Happens When the Endosymbionts "Bug Out"? [2]
  • Introduction
  • Background
  • Methods
  • Insect rearing
  • S. pierantonius DNA quantification with qPCR
  • Fluorescence microscopy
  • Electron microcopy
  • Cuticle coloration
  • Results
  • Subchapter 15.3 - Parvovirus: Hijacking Endocytosis [3]
  • Introduction
  • Background
  • Methods
  • Cells and viruses
  • Lysosomal drugs
  • Microscopy
  • Assay of CPV infectivity
  • Dextran particle release assay
  • Results
  • References
  • Subject Index
  • Back Cover

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