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[资源] Current Protocols in Cell Biology (Wiley, 2004) .pdf

Current Protocols in Cell Biology (Wiley, 2004)
Chapter 1 Cell Culture
Introduction HTML PDF (Size 70KB)
Unit 1.1 Basic Techniques for Mammalian Cell Tissue Culture HTML PDF (Size 170KB)
Unit 1.2 Media for Culture of Mammalian Cells HTML PDF (Size 224KB)
Unit 1.3 Aseptic Technique for Cell Culture HTML PDF (Size 172KB)
Unit 1.4 Sterilization and Filtration HTML PDF (Size 230KB)
Unit 1.5 Assessing and Controlling Microbial Contamination in Cell Cultures HTML PDF (Size 463KB)
Unit 1.6 Media and Culture of Yeast HTML PDF (Size 176KB)
Unit 1.7 BY-2 Cells: Culture and Transformation for Live Cell Imaging HTML PDF (Size 332KB)
Chapter 2 Preparation and Isolation of Cells
Introduction HTML PDF (Size 63KB)
Unit 2.1 Establishment of Fibroblast Cultures HTML PDF (Size 706KB)
Unit 2.2 Preparation and Culture of Human Lymphocytes HTML PDF (Size 165KB)
Unit 2.3 Preparation of Endothelial Cells HTML PDF (Size 270KB)
Unit 2.4 Generation of Continuously Growing B Cell Lines by Epstein-Barr Virus Transformation HTML PDF (Size
100KB)
Unit 2.5 Laser Capture Microdissection HTML PDF (Size 298KB)
Unit 2.6 Preparation of Human Epidermal Keratinocyte Cultures HTML PDF (Size 260KB)
Chapter 3 Subcellular Fractionation and Isolation of Organelles
Introduction HTML PDF (Size 34KB)
Unit 3.1 Overview of Cell Fractionation HTML PDF (Size 193KB)
Unit 3.2 Isolation of Rat Hepatocyte Plasma Membrane Sheets and Plasma Membrane Domains HTML PDF (Size 349KB)
Unit 3.3 Isolation of Mitochondria from Tissues and Cells by Differential Centrifugation HTML PDF (Size 187KB)
Unit 3.4 Purification of a Crude Mitochondrial Fraction by Density-Gradient Centrifugation HTML PDF (Size 269KB)
Unit 3.5 Isolation of Peroxisomes from Tissues and Cells by Differential and Density Gradient
Centrifugation HTML PDF (Size 206KB)
Unit 3.6 Isolation of Lysosomes from Tissues and Cells by Differential and Density Gradient
Centrifugation HTML PDF (Size 214KB)
Unit 3.7 Overview of Subcellular Fractionation Procedures for the Yeast Saccharomyces cerevisiae HTML PDF (Size
273KB)
Unit 3.8 Isolation of Subcellular Fractions from the Yeast Saccharomyces cerevisiae HTML PDF (Size 503KB)
Unit 3.9 Isolation of Golgi Membranes from Tissues and Cells by Differential and Density Gradient
Centrifugation HTML PDF (Size 221KB)
Unit 3.10 Isolation of Nuclei and Nuclear Membranes From Animal Tissues HTML PDF (Size 174KB)
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Unit 3.11 Free-Flow Electrophoretic Analysis of Endosome Subpopulations of Rat Hepatocytes HTML PDF (Size 390KB)
Chapter 4 Microscopy
Introduction HTML PDF (Size 87KB)
Unit 4.1 Proper Alignment and Adjustment of the Light Microscope HTML PDF (Size 929KB)
Unit 4.2 Fluorescence Microscopy HTML PDF (Size 178KB)
Unit 4.3 Immunofluorescence Staining HTML PDF (Size 228KB)
Unit 4.4 Fluorescent Staining of Subcellular Organelles: ER, Golgi Complex, and Mitochondria HTML PDF (Size 503KB)
Unit 4.5 Basic Confocal Microscopy HTML PDF (Size 358KB)
Unit 4.6 Immunoperoxidase Methods for Localization of Antigens in Cultured Cells and Tissues HTML PDF (Size 439KB)
Unit 4.7 Cryo-Immunogold Electron Microscopy HTML PDF (Size 497KB)
Unit 4.8 Correlative Video Light/Electron Microscopy HTML PDF (Size 407KB)
Unit 4.9 Polarization Microscopy HTML PDF (Size 1367KB)
Unit 4.10 Fluorescent Speckle Microscopy (FSM) of Microtubules and Actin in Living Cells HTML PDF (Size 562KB)
Unit 4.11 Two-Photon Excitation Microscopy for the Study of Living Cells and Tissues HTML PDF (Size 1421KB)
Unit 4.12 Total Internal Reflection Fluorescence Microscopy for High-Resolution Imaging of Cell-Surface
Events HTML PDF (Size 430KB)
Unit 4.13 Fluorescent Labeling of Yeast HTML PDF (Size 524KB)
Unit 4A Organelle Atlas: Appendix to Chapter 4 HTML PDF (Size 2384KB)
Chapter 5 Characterization of Cellular Proteins
Introduction HTML PDF (Size 75KB)
Unit 5.1 Overview of the Physical State of Proteins Within Cells HTML PDF (Size 187KB)
Unit 5.2 Determining the Topology of an Integral Membrane Protein HTML PDF (Size 337KB)
Unit 5.3 Determination of Molecular Size by Zonal Sedimentation Analysis on Sucrose Density
Gradients HTML PDF (Size 311KB)
Unit 5.4 Analysis of the Association of Proteins with Membranes HTML PDF (Size 230KB)
Unit 5.5 Determination of Molecular Size by Size-Exclusion Chromatography (Gel Filtration) HTML PDF (Size 233KB)
Unit 5.6 Identification of Proteins in Complex Mixtures Using Liquid Chromatography and Mass
Spectrometry HTML PDF (Size 308KB)
Chapter 6 Electrophoresis and Immunoblotting
Introduction HTML PDF (Size 48KB)
Unit 6.1 One-Dimensional SDS Gel Electrophoresis of Proteins HTML PDF (Size 404KB)
Unit 6.2 Immunoblotting and Immunodetection HTML PDF (Size 245KB)
Unit 6.3 Detection and Quantitation of Radiolabeled Proteins in Gels and Blots HTML PDF (Size 134KB)
Unit 6.4 Two-Dimensional Gel Electrophoresis HTML PDF (Size 290KB)
Unit 6.5 One-Dimensional Electrophoresis Using Nondenaturing Conditions HTML PDF (Size 208KB)
Unit 6.6 Staining Proteins in Gels HTML PDF (Size 209KB)
Unit 6.7 Agarose Gel Electrophoresis of Proteins HTML PDF (Size 187KB)
Unit 6.8 Fluorescence Detection of Glycoproteins in Gels and on Electroblots HTML PDF (Size 218KB)
Unit 6.9 Digital Electrophoresis Analysis HTML PDF (Size 262KB)
Chapter 7 Protein Labeling and Immunoprecipitation
Introduction HTML PDF (Size 52KB)
Unit 7.1 Metabolic Labeling with Amino Acids HTML PDF (Size 147KB)
Unit 7.2 Immunoprecipitation HTML PDF (Size 271KB)
Unit 7.3 Metabolic Labeling with Sulfate HTML PDF (Size 98KB)
Unit 7.4 Metabolic Labeling with Fatty Acids HTML PDF (Size 175KB)
Unit 7.5 Metabolic Labeling of Prenyl and Carboxyl-Methyl Groups HTML PDF (Size 143KB)
Unit 7.6 Metabolic Labeling and Immunoprecipitation of Yeast Proteins HTML PDF (Size 111KB)
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Unit 7.7 Metabolic Labeling and Immunoprecipitation of Drosophila Proteins HTML PDF (Size 91KB)
Unit 7.8 Metabolic Labeling of Glycoproteins with Radioactive Sugars HTML PDF (Size 149KB)
Unit 7.9 Analysis of Oxidative Modification of Proteins HTML PDF (Size 239KB)
Unit 7.10 Radioiodination of Cellular Proteins HTML PDF (Size 156KB)
Chapter 8 Cell Cycle Analysis
Introduction HTML PDF (Size 40KB)
Unit 8.1 Overview of the Cell Cycle HTML PDF (Size 153KB)
Unit 8.2 Assays for CDK Activity and DNA Replication in the Cell Cycle HTML PDF (Size 168KB)
Unit 8.3 Methods for Synchronizing Cells at Specific Stages of the Cell Cycle HTML PDF (Size 213KB)
Unit 8.4 Determining Cell Cycle Stages by Flow Cytometry HTML PDF (Size 278KB)
Unit 8.5 Centrifugal Elutriation to Obtain Synchronous Populations of Cells HTML PDF (Size 246KB)
Chapter 9 Cell Adhesion
Introduction HTML PDF (Size 149KB)
Unit 9.1 Cell-Substrate Adhesion Assays HTML PDF (Size 139KB)
Unit 9.2 Quantitative Measurement of Cell Adhesion Using Centrifugal Force HTML PDF (Size 183KB)
Unit 9.3 Cadherin-Dependent Cell-Cell Adhesion HTML PDF (Size 161KB)
Unit 9.4 Analyzing Integrin-Dependent Adhesion HTML PDF (Size 273KB)
Unit 9.5 Analysis of Cell-Cell Contact Mediated by Ig Superfamily Cell Adhesion Molecules HTML PDF (Size 819KB)
Unit 9.6 Measurement of Adhesion Under Flow Conditions HTML PDF (Size 241KB)
Chapter 10 Extracellular Matrix
Introduction HTML PDF (Size 97KB)
Unit 10.1 Overview of Extracellular Matrix HTML PDF (Size 278KB)
Unit 10.2 Preparation of Basement Membrane Components from EHS Tumors HTML PDF (Size 112KB)
Unit 10.3 Preparation of Gelled Substrates HTML PDF (Size 142KB)
Unit 10.4 Preparation of Extracellular Matrices Produced by Cultured Corneal Endothelial and PF-HR9 Endodermal
Cells HTML PDF (Size 282KB)
Unit 10.5 Purification of Fibronectin HTML PDF (Size 131KB)
Unit 10.6 Purification of Vitronectin HTML PDF (Size 90KB)
Unit 10.7 Proteoglycan Isolation and Analysis HTML PDF (Size 166KB)
Unit 10.8 Matrix Metalloproteinases HTML PDF (Size 836KB)
Unit 10.9 Preparation of Extracellular Matrices Produced by Cultured Fibroblasts HTML PDF (Size 388KB)
Unit 10.10 Purification and Analysis of Thrombospondin-1 HTML PDF (Size 223KB)
Unit 10.11 Purification of SPARC/Osteonectin HTML PDF (Size 266KB)
Chapter 11 In Vitro Reconstitution
Introduction HTML PDF (Size 87KB)
Unit 11.1 Overview of Eukaryotic In Vitro Translation and Expression Systems HTML PDF (Size 148KB)
Unit 11.2 In Vitro Translation HTML PDF (Size 206KB)
Unit 11.3 In Vitro Analysis of Endoplasmic-Reticulum-to-Golgi Transport in Mammalian Cells HTML PDF (Size 275KB)
Unit 11.4 Cotranslational Translocation of Proteins into Canine Rough Microsomes HTML PDF (Size 197KB)
Unit 11.5 In Vitro Analysis of SV40 DNA Replication HTML PDF (Size 164KB)
Unit 11.6 In Vitro Transcription HTML PDF (Size 211KB)
Unit 11.7 Nuclear Import in Digitonin-Permeabilized Cells HTML PDF (Size 439KB)
Unit 11.8 In Vitro Translation Using HeLa Extract HTML PDF (Size 144KB)
Unit 11.9 Analysis of Eukaryotic Translation in Purified and Semipurified Systems HTML PDF (Size 292KB)
Unit 11.10 Preparation and Use of Interphase Xenopus Egg Extracts HTML PDF (Size 332KB)
Unit 11.11 Analysis of the Cell Cycle Using Xenopus Egg Extracts HTML PDF (Size 215KB)
Unit 11.12 Analysis of Apoptosis Using Xenopus Egg Extracts HTML PDF (Size 155KB)
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Unit 11.13 Mitotic Spindle Assembly In Vitro HTML PDF (Size 329KB)
Unit 11.14 Analysis of RNA Export Using Xenopus Oocytes HTML PDF (Size 208KB)
Unit 11.15 In Vitro Analysis of Peroxisomal Protein Import HTML PDF (Size 178KB)
Unit 11.16 In Vitro Analysis of Chloroplast Protein Import HTML PDF (Size 234KB)
Unit 11.17 In Vitro RNA Splicing in Mammalian Cell Extracts HTML PDF (Size 315KB)
Chapter 12 Cell Motility
Introduction HTML PDF (Size 30KB)
Unit 12.1 Chemotaxis Assays for Eukaryotic Cells HTML PDF (Size 871KB)
Unit 12.2 Invasion Assays HTML PDF (Size 158KB)
Unit 12.3 Cell Traction HTML PDF (Size 440KB)
Unit 12.4 Cell Wound Assays HTML PDF (Size 477KB)
Unit 12.5 Dictyostelium Cell Dynamics HTML PDF (Size 629KB)
Unit 12.6 Optical Microscopy—Based Migration Assay for Human Neutrophils HTML PDF (Size 547KB)
Unit 12.7 Actin-Based Motility Assay HTML PDF (Size 259KB)
Chapter 13 Organelle Motility
Introduction HTML PDF (Size 62KB)
Unit 13.1 Microtubule/Organelle Motility Assays HTML PDF (Size 263KB)
Unit 13.2 In Vitro Motility Assay to Study Translocation of Actin by Myosin HTML PDF (Size 118KB)
Unit 13.3 Organelle Motility in Plant Cells: Imaging Golgi and ER Dynamics with GFP HTML PDF (Size 394KB)
Unit 13.4 Movement of Nuclei HTML PDF (Size 450KB)
Unit 13.5 Measuring Dynamics of Nuclear Proteins by Photobleaching HTML PDF (Size 248KB)
Chapter 14 Signal Transduction: Protein Phosphorylation
Introduction HTML PDF (Size 42KB)
Unit 14.1 Overview of Protein Phosphorylation HTML PDF (Size 83KB)
Unit 14.2 Immunological Detection of Phosphorylation HTML PDF (Size 234KB)
Unit 14.3 Detection of MAP Kinase Signaling HTML PDF (Size 298KB)
Unit 14.4 Labeling Cultured Cells with 32Pi and Preparing Cell Lysates for Immunoprecipitation HTML PDF (Size 120KB)
Unit 14.5 Phosphoamino Acid Analysis HTML PDF (Size 143KB)
Unit 14.6 Determination of Akt/PKB Signaling HTML PDF (Size 271KB)
Chapter 15 Protein Trafficking
Introduction HTML PDF (Size 89KB)
Unit 15.1 Overview of Protein Trafficking in the Secretory and Endocytic Pathways HTML PDF (Size 213KB)
Unit 15.2 Use of Glycosidases to Study Protein Trafficking HTML PDF (Size 317KB)
Unit 15.3 Endocytosis: Biochemical Analyses HTML PDF (Size 190KB)
Unit 15.4 Determining Protein Transport to the Plasma Membrane HTML PDF (Size 149KB)
Unit 15.5 Analysis of Membrane Traffic in Polarized Epithelial Cells HTML PDF (Size 201KB)
Unit 15.6 Analysis of Protein Folding and Oxidation in the Endoplasmic Reticulum HTML PDF (Size 375KB)
Unit 15.7 Measurements of Phagocytosis and Phagosomal Maturation HTML PDF (Size 383KB)
Chapter 16 Antibodies as Cell Biological Tools
Introduction HTML PDF (Size 45KB)
Unit 16.1 Production of Monoclonal Antibodies HTML PDF (Size 189KB)
Unit 16.2 Production of Polyclonal Antisera HTML PDF (Size 133KB)
Unit 16.3 Purification of Immunoglobulin G HTML PDF (Size 170KB)
Unit 16.4 Fragmentation of Immunoglobulin G HTML PDF (Size 131KB)
Unit 16.5 Antibody Conjugates for Cell Biology HTML PDF (Size 253KB)
Unit 16.6 Production of Antibodies That Recognize Specific Tyrosine-Phosphorylated Peptides HTML PDF (Size 205KB)
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Chapter 17 Macromolecular Interactions in Cells
Introduction HTML PDF (Size 29KB)
Unit 17.1 Imaging Protein-Protein Interactions by Fluorescence Resonance Energy Transfer (FRET)
Microscopy HTML PDF (Size 342KB)
Unit 17.2 Identification of Protein Interactions by Far Western Analysis HTML PDF (Size 196KB)
Unit 17.3 Interaction Trap/Two-Hybrid System to Identify Interacting Proteins HTML PDF (Size 463KB)
Unit 17.4 Mapping Protein-Protein Interactions with Phage-Displayed Combinatorial Peptide Libraries HTML PDF (Size
196KB)
Unit 17.5 Protein-Protein Interactions Identified by Pull-Down Experiments and Mass Spectrometry HTML PDF (Size
679KB)
Unit 17.6 Measuring Protein Interactions by Optical Biosensors HTML PDF (Size 356KB)
Unit 17.7 Chromatin Immunoprecipitation for Determining the Association of Proteins with Specific Genomic Sequences In
Vivo HTML PDF (Size 240KB)
Unit 17.8 Isothermal Titration Calorimetry HTML PDF (Size 486KB)
Chapter 18 Cellular Aging and Death
Introduction HTML PDF (Size 81KB)
Unit 18.1 Current Concepts in Cell Death HTML PDF (Size 306KB)
Unit 18.2 Analysis of Caspase Activation During Apoptosis HTML PDF (Size 380KB)
Unit 18.3 Assessment of Apoptosis and Necrosis by DNA Fragmentation and Morphological Criteria HTML PDF (Size
357KB)
Unit 18.4 Quantitative Fluorescence In Situ Hybridization (Q-FISH) HTML PDF (Size 405KB)
Unit 18.5 Analysis of Mitochondrial Dysfunction During Cell Death HTML PDF (Size 360KB)
Unit 18.6 Analysis of Telomeres and Telomerase HTML PDF (Size 647KB)
Unit 18.7 Nonisotopic Methods for Determination of Poly(ADP-Ribose) Levels and Detection of Poly(ADP-Ribose)
Polymerase HTML PDF (Size 718KB)
Unit 18.8 Flow Cytometry of Apoptosis HTML PDF (Size 530KB)
Chapter 19 Whole Organism and Tissue Analysis
Introduction HTML PDF (Size 82KB)
Unit 19.1 Overview of Metastasis Assays HTML PDF (Size 217KB)
Unit 19.2 Tail Vein Assay of Cancer Metastasis HTML PDF (Size 186KB)
Unit 19.3 Microanalysis of Gene Expression in Tissues Using T7-SAGE: Serial Analysis of Gene Expression After High-
Fidelity T7-Based RNA Amplification HTML PDF (Size 349KB)
Unit 19.4 SAGE Analysis from 1 μg of Total RNA HTML PDF (Size 262KB)
Unit 19.5 The Chick Chorioallantoic Membrane as an In Vivo Angiogenesis Model HTML PDF (Size 218KB)
Unit 19.6 Experimental Metastasis Assays in the Chick Embryo HTML PDF (Size 568KB)
Chapter 20 Expression and Introduction of Macromolecules into Cells
Introduction HTML PDF (Size 64KB)
Unit 20.1 Direct Introduction of Molecules into Cells HTML PDF (Size 149KB)
Unit 20.2 Protein Transduction: Generation of Full-Length Transducible Proteins Using the TAT
System HTML PDF (Size 543KB)
Unit 20.3 Calcium Phosphate Transfection HTML PDF (Size 184KB)
Unit 20.4 Transfection Using DEAE-Dextran HTML PDF (Size 199KB)
Unit 20.5 Transfection by Electroporation HTML PDF (Size 147KB)
Unit 20.6 Transfection of Cultured Eukaryotic Cells Using Cationic Lipid Reagents HTML PDF (Size 409KB)
Unit 20.7 Optimization of Transfection HTML PDF (Size 149KB)
Chapter 21 Fluorescent Protein Technology
Introduction HTML PDF (Size 28KB)
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Unit 21.1 Measuring Protein Mobility by Photobleaching GFP Chimeras in Living Cells HTML PDF (Size 547KB)
Unit 21.2 Fluorescence Localization After Photobleaching (FLAP) HTML PDF (Size 896KB)
Chapter 22 Cell Biology of Chromosomes and Nuclei
Introduction HTML PDF (Size 28KB)
Unit 22.1 Overview of Cytogenetic Chromosome Analysis HTML PDF (Size 104KB)
Unit 22.2 Preparation of Cytogenetic Specimens from Tissue Samples HTML PDF (Size 186KB)
Unit 22.3 Traditional Banding of Chromosomes for Cytogenetic Analysis HTML PDF (Size 453KB)
Unit 22.4 Fluorescence In Situ Hybridization (FISH) HTML PDF (Size 708KB)
Unit 22.5 Multi-Color FISH Techniques HTML PDF (Size 730KB)
Appendix 1 Useful Information and Data
1A Useful Measurements and Data HTML PDF (Size 46KB)
1B Compendium of Drugs Commonly Used in Cell Biology Research HTML PDF (Size 370KB)
1C Identification of Motifs in Protein Sequences HTML PDF (Size 205KB)
1D Safe Use of Radioisotopes HTML PDF (Size 179KB)
1E Absorption and Emission Maxima for Common Fluorophores HTML PDF (Size 101KB)
1F Importing Biological Materials HTML PDF (Size 88KB)
1G Centrifuges and Rotors HTML PDF (Size 83KB)
1H Internet Basics for Biologists HTML PDF (Size 164KB)
Appendix 2 Laboratory Stock Solutions and Equipment
2A Common Stock Solutions, Buffers, and Media HTML PDF (Size 181KB)
2B Medium Formulations HTML PDF (Size 113KB)
2C Standard Laboratory Equipment HTML PDF (Size 89KB)
Appendix 3 Commonly Used Techniques
3A Molecular Biology Techniques HTML PDF (Size 38KB)
3B Spectrophotometric Determination of Protein Concentration HTML PDF (Size 85KB)
3C Dialysis and Concentration of Protein Solutions HTML PDF (Size 165KB)
3D Quantification of DNA and RNA with Absorption and Fluorescence Spectroscopy HTML PDF (Size 161KB)
3E Silanizing Glassware HTML PDF (Size 35KB)
3F Enzymatic Amplification of DNA by PCR: Standard Procedures and Optimization HTML PDF (Size 153KB)
3G Micro RT-PCR HTML PDF (Size 72KB)
3H The Colorimetric Detection and Quantitation of Total Protein HTML PDF (Size 238KB)
SUPPLIERS APPENDIX
Selected Suppliers of Reagents and Equipment HTML PDF (Size 179KB)

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