Subcellular fractionation: a practical approach
Gespeichert in:
| Format: | Buch |
|---|---|
| Sprache: | English |
| Veröffentlicht: |
Oxford [u.a.]
IRL Press
1997
|
| Schriftenreihe: | The practical approach series
173 |
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | XX, 339 S. Ill., graph. Darst. |
| ISBN: | 0199634955 0199634947 |
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Datensatz im Suchindex
| _version_ | 1804125794858958848 |
|---|---|
| adam_text | SUBCELLULAR FRACTIONATION A PRACTICAL APPROACH EDITED BY J. M. GRAHAM
JOHN GRAHAM RESEARCH CONSULTANCY, SCHOOL OF BIOMOLECULAR SCIENCES,
LIVERPOOL JOHN MOORES UNIVERSITY, LIVERPOOL L3 3AF, UK AND D. RICKWOOD
DEPARTMENT OF BIOLOGY, UNIVERSITY OF ESSEX, COLCHESTER CO4 3SQ, UK
TECHNISCHE UNIVERSITST DARMSTADT FACHBEREICH 10 * BIOLQGIE * B I B ! I O
T H E K * SOHNLTTSPAHNSTRABS 10 D-6 4 2 8 7 DARMSTADT -AT- OXFORD
UNIVERSITY PRESS OXFORD NEW YORK TOKYO CONTENTS LIST OF CONTRIBUTORS
XVII ABBREVIATIONS XIX 1. HOMOGENIZATION OF TISSUES AND CELLS 1 JOHN M.
GRAHAM 1. INTRODUCTION 1 2. AIMS OF THE HOMOGENIZATION PROCEDURE 1 3.
INFLUENCE OF SAMPLE TYPE 3 4. HOMOGENIZATION MEDIA 4 5. METHODS OF
HOMOGENIZATION 6 TYPE 1 HOMOGENIZERS 7 TYPE 2 HOMOGENIZERS 10 6.
HOMOGENIZATION OF TISSUES AND CELLS 14 MAMMALIAN LIVER 14 BRAIN 17
MUSCLE 18 MAMMALIAN TISSUE CULTURE CELLS 19 PLANT ORGANELLES 22 YEAST 23
OTHER FUNGI AND ALGAE 24 TRYPANOSOMES 25 BACTERIA 26 REFERENCES 28 2.
ISOLATION OF SUBCELLULAR FRACTIONS 31 RICHARD H. HINTON AND BARBARA M.
MULLOCK 1. INTRODUCTION 31 2. COMPOSITION OF A TISSUE HOMOGENATE 34 3.
PROPERTIES OF CELL ORGANELLES 38 FACTORS AFFECTING ORGANELLE DENSITY AND
SIZE 38 4. CENTRIFUGAL METHODS FOR THE SEPARATION OF ORGANELLES 40
SEPARATION BY SIZE 41 SEPARATION BY DENSITY 43 DENSITY PERTURBATION 49
CONTENTS 5. NON-CENTRIFUGAL PROCEDURES 50 IMMUNOISOLATION 50 SEPARATION
BY ELECTROPHORESIS 51 6. IDENTIFICATION OF SEPARATED MATERIAL 51 MARKER
ENZYMES 51 INTRODUCED MARKERS FOR ENDOCYTIC AND EXOCYTIC PATHWAYS 54
CHARACTERISTIC NON-ENZYMATIC PROTEINS 54 7. ASSESSMENT OF THE PURITY OF
FRACTIONS 55 PURITY AND PURIFICATION 56 PROBLEMS FROM CELL HETEROGENEITY
WITHIN TISSUES 56 PROBLEMS ARISING FROM ORGANELLE FRAGMENTATION 57
MISSORTING IN THE EXOCYTIC AND ENDOCYTIC PATHWAYS 57 8. FRACTIONATION
PROBLEMS 58 NO SEPARATION 58 AGGREGATION FOLLOWING RESUSPENSION OF A
FRACTION 58 POOR RECOVERY OF MARKERS 58 DAMAGE TO CELL STRUCTURES 60 9.
A SYSTEMATIC APPROACH TO CELL FRACTIONATION 60 PRELIMINARY STUDIES 61
DETERMINATION OF THE PROPERTIES OF COMPONENTS OF THE HOMOGENATE 61
METHOD DEVELOPMENT 63 SIMPLIFICATION OF THE SEPARATION 65 REFERENCES 69
3. ISOLATION AND CHARACTERIZATION OF NUCLEI AND NUCLEAR SUBTRACTIONS 71
DAVID RICKWOOD, ANTHEA MESSENT, AND DIPAK PATEL 1. INTRODUCTION 71 2.
METHODS OF PREPARING PURIFIED NUCLEI 71 TYPES OF CELLS AND TISSUE
SAMPLES 71 HOMOGENIZATION MEDIA 72 HOMOGENIZATION METHODS 73
CENTRIFUGATION CONDITIONS 73 ASSAYS OF NUCLEAR PURITY 77 3. METHODS FOR
PURIFYING METAPHASE CHROMOSOMES 78 4. ISOLATION OF NUCLEAR SUBTRACTIONS
80 PREPARATION OF NUCLEOLI 80 PREPARATION OF NUCLEAR MEMBRANES 81
ISOLATION OF NUCLEAR MATRIX 84 PREPARATION OF NUCLEOIDS 85 5. ISOLATION
OF NUCLEOPROTEIN COMPLEXES 86 CONTENTS ISOLATION OF POLYNUCLEOSOMES OF
CHROMATIN 86 RIBONUCLEOPROTEINS 88 6. ISOLATION OF NUCLEAR
MACROMOLECULES 89 ISOLATION OF NUCLEAR PROTEINS 89 ISOLATION OF NUCLEAR
RNA 91 ISOLATION OF DNA 97 7. FUNCTIONAL ASSAYS OF NUCLEI 99 ANALYSIS OF
DNA-BINDING PROTEINS 99 TRANSCRIPTION ASSAYS 101 REFERENCES 103 4.
SUBCELLULAR FRACTIONATION OF MITOCHONDRIA 107 /. E. RICE AND J. G.
LINDSAY 1. INTRODUCTION 107 2. PURIFICATION OF MITOCHONDRIA FROM VARIOUS
EUKARYOTIC SOURCES 108 INTRODUCTION 108 PROTOCOLS FOR PURIFICATION OF
MITOCHONDRIA FROM SEVERAL EUKARYOTIC SOURCES . 112 FURTHER PURIFICATION
OF MITOCHONDRIAL FRACTIONS 119 3. DETERMINATION OF MITOCHONDRIAL PURITY
119 INTRODUCTION 119 USE OF THE OXYGEN ELECTRODE TO DETERMINE
MITOCHONDRIAL INTEGRITY 120 DETERMINATION OF THE INTEGRITY OF THE
MITOCHONDRIAL OUTER MEMBRANE 127 GLUCOSE HEXOKINASE TRAP METHOD FOR
ESTIMATION OF P:O RATIOS 128 4. SUBFRACTIONATION OF MITOCHONDRIA 130
PREPARATION OF SUBMITOCHONDRIAL PARTICLES BY SONICATION 130 PREPARATION
OF MITOPLASTS USING DIGITONIN 131 SEPARATION OF RIGHT SIDE OUT AND
INSIDE OUT BY SUBMITOCHONDRIAL PARTICLES BY AFFINITY CHROMATOGRAPHY ON
CNBR-CYTOCHROME C. COUPLED SEPHAROSE 133 ASSAYING OF MITOCHONDRIAL
MARKER ENZYMES 135 5. MEDICAL ASPECTS OF MITOCHONDRIAL RESEARCH 136
MITOCHONDRIAL BIOGENESIS 136 OXIDATIVE PHOSPHORYLATION AND THE ELECTRON
TRANSPORT CHAIN 137 MITOCHONDRIAL MYOPATHIES 138 TECHNIQUES APPLIED IN
THE INVESTIGATION OF MITOCHONDRIAL ABNORMALITIES 139 6. CONCLUDING
REMARKS 141 REFERENCES 142 XI CONTENTS 5. ISOLATION AND CHARACTERIZATION
OF PEROXISOMES 143 A. VOLKL, E. BAUMGART, AND H. D. FAHIMI 1.
INTRODUCTION 143 2. PREPARATION AND PURIFICATION OF RAT HEPATIC PO 144
GENERAL CONSIDERATIONS 144 EXPERIMENTAL DESIGN 145 ANALYSIS OF RESULTS -
150 3. SUBFRACTIONATION OF PURIFIED RAT HEPATIC PEROXISOMES 153
EXTRACTION OF SOLUBLE MATRIX PROTEINS 153 EXTRACTION OF CORE-BOUND URATE
OXIDASE 153 ISOLATION OF INTEGRAL MEMBRANE PROTEINS OF PO 153 4.
COMPARATIVE CHARACTERIZATION OF ISOLATED HEPATIC PEROXISOMES FROM
DIFFERENT SPECIES 155 ENZYME ACTIVITIES OF PO FRACTIONS 155
IMMUNOBLOTTING OF PO FRACTIONS 157 5. EMBEDDING OF PO FRACTIONS FOR
ELECTRON MICROSCOPY (EM) 158 EQUIPMENT AND SOLUTIONS 158 PROCESSING OF
PO FRACTIONS FOR EM 161 RESULTS 165 ACKNOWLEDGEMENTS 165 REFERENCES 165
6. LYSOSOMES AND ENDOCYTOSIS 169 TOR GJ0EN, TROND OLAV BERG, AND TROND
BERG 1. CURRENT MODELS OF ENDOCYTOSIS 169 2. ANALYSIS OF ENDOCYTIC
UPTAKE OF LIGANDS AND PINOCYTOSIS MARKERS 170 RECEPTOR-MEDIATED
ENDOCYTOSIS 170 FLUID PHASE ENDOCYTOSIS 176 3. ENDOCYTOSIS OF PLASMA
MEMBRANE COMPONENTS 177 4. ANALYSIS OF INTRACELLULAR TRANSPORT OF
LIGANDS/MARKERS AND ENDOCYTIC RECEPTORS BY MEANS OF SUBCELLULAR
FRACTIONATION 178 DIFFERENTIAL CENTRIFUGATION 178 ISOPYCNIC
CENTRIFUGATION IN GRADIENTS OF SUCROSE, NYCODENZ, OR PERCOLL 180
CONTINUOUS FLOW ELECTROPHORESIS (CFE) 185 5. LABELLING OF LIGANDS AND
MARKERS 185 USE OF [ 125 I] 185 XII CONTENTS USE OF RESIDUALIZING LABELS
186 LABELLING OF RECEPTORS 188 6. MARKERS FOR ORGANELLES INVOLVED IN
ENDOCYTOSIS 189 THE PLASMA MEMBRANE 189 ENDOSOMES 191 THE LYSOSOMES 191
7. INDUCTION OF DENSITY SHIFT OF COMPONENTS OF THE ENDOCYTIC PATHWAY TO
DETERMINE A POSSIBLE CO-LOCALIZATION OF LIGAND AND OTHER MARKERS 192
DENSITY SHIFT INDUCED IN ENDOSOMES THAT CONTAIN LIGAND IN COMPLEX WITH
HORSE-RADISH PEROXIDASE OR COLLOIDAL GOLD 193 8. USE OF SPECIFIC
PERTURBANTS TO ACCUMULATE LIGANDS AND/OR RECEPTORS AT DEFINED
INTRACELLULAR SITES 196 9. USE OF LYSOSOMAL ENZYME SUBSTRATES TO DISRUPT
LYSOSOMES SELECTIVELY 198 10. CONCLUDING REMARKS 200 REFERENCES 200 7.
THE MEMBRANES OF THE SECRETORY AND EXOCYTIC PATHWAYS 205 JOHN M. GRAHAM
1. INTRODUCTION 205 GRADIENT MEDIA 205 2. ISOLATION OF PLASMA MEMBRANE,
ENDOPLASMIC RETICULUM, AND GOLGI FROM RAT LIVER 208 PLASMA MEMBRANE 208
SEPARATION OF PLASMA MEMBRANE DOMAINS 211 ENDOPLASMIC RETICULUM (ER) 212
GOLGI MEMBRANES 214 ISOLATION OF MEMBRANES BY IMMUNOAFFINITY 221 DENSITY
PERTURBATION METHODS 224 3. THE SECRETORY PATHWAY 225 RESOLUTION OF
GOLGI DOMAINS 225 IDENTIFICATION OF VESICLES INVOLVED IN TRANSPORT TO
THE GOLGI 229 IDENTIFICATION OF VESICLES INVOLVED IN TRANSPORT FROM THE
GOLGI 231 IN VITRO SYSTEMS TO STUDY TRANSFER OF MOLECULES BETWEEN
MEMBRANES 233 SORTING OF PROTEINS DESTINED FOR SPECIFIC PLASMA MEMBRANE
DOMAINS 234 SECRETORY PATHWAYS IN YEAST 236 GEL FILTRATION 238
REFERENCES 239 XIII CONTENTS 8. ISOLATION AND PURIFICATION OF
FUNCTIONALLY INTACT MITOCHONDRIA AND CHLOROPLASTS FROM PLANT CELLS 243
ANTHONY L. MOORE AND DAVID G. WHITEHOUSE 1. INTRODUCTION 243 2.
MITOCHONDRIA 243 GENERAL ISOLATION PRINCIPLES 245 3. CHLOROPLASTS 249
GENERAL ISOLATION PRINCIPLES 251 4. ASSAYS 260 MITOCHONDRIA 260
CHLOROPLASTS 265 ACKNOWLEDGEMENTS 268 REFERENCES 268 9. RIBOSOMES AND
POLYSOMES 271 ULRICH A. BOMMER, NILS BURKHARDT, RALF JIINEMANN,
CHRISTIAN M. T. SPAHN, FRANCISCO J. TRIANA-ALONSO, AND KNUD H. NIERHAUS
1. INTRODUCTION 271 2. ISOLATION OF RIBOSOMES AND POLYSOMES 271
PROKARYOTIC RIBOSOMES 271 EUKARYOTIC RIBOSOMES 280 3. ASSAY SYSTEMS 287
PROTEIN SYNTHESIS SYSTEMS 287 TESTING PARTIAL REACTIONS OF THE RIBOSOME
CYCLE 292 ACKNOWLEDGEMENTS 300 REFERENCES 300 10. ELECTRON MICROSCOPY OF
ORGANELLES 303 NIGEL JAMES 1. INTRODUCTION 303 2. FIXATION 304
IMPORTANCE OF FIXATION 304 CHEMICAL FIXATION 305 OSMOLARITY OF FIXATIVE
SOLUTIONS 307 XIV CONTENTS TEMPERATURE, PH, AND OTHER FACTORS 308
CRYOFIXATION 310 CRYOULTRAMICROTOMY 311 FREEZE-FRACTURE PREPARATIONS 312
3. EMBEDDING 312 HARDENERS, MODIFIERS, AND CATALYSTS 314 4. MICROTOMY
314 5. STAINING 316 THE ELECTRON MICROSCOPE FAMILY . 316 IMAGE CONTRAST
316 STRUCTURAL STAINING 318 ANTIBODY STAINING 319 COLLOIDAL GOLD 319
ENZYME CYTOCHEMISTRY 321 HISTOCHEMISTRY 323 NEGATIVE STAINING 325 6.
FURTHER READING 326 REFERENCES 327 AL. CORE LIST OF SUPPLIERS 329 INDEX
333 XV
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| id | DE-604.BV011289955 |
| illustrated | Illustrated |
| indexdate | 2024-07-09T18:07:14Z |
| institution | BVB |
| isbn | 0199634955 0199634947 |
| language | English |
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| publisher | IRL Press |
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| series | The practical approach series |
| series2 | The practical approach series |
| spelling | Subcellular fractionation a practical approach ed. by J. M. Graham ... Oxford [u.a.] IRL Press 1997 XX, 339 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier The practical approach series 173 Cellen (biologie) gtt Fractionering gtt Weefsel gtt Subcellular Fractions analysis Subcellular Fractions methods Subcellular fractionation Handbooks, manuals, etc Subcellular fractionation Laboratory manuals Zelle (DE-588)4067537-3 gnd rswk-swf Kompartiment (DE-588)4426443-4 gnd rswk-swf Fraktionierung (DE-588)4018056-6 gnd rswk-swf Methode (DE-588)4038971-6 gnd rswk-swf Zelle (DE-588)4067537-3 s Kompartiment (DE-588)4426443-4 s Fraktionierung (DE-588)4018056-6 s DE-604 Methode (DE-588)4038971-6 s Graham, John M. Sonstige oth The practical approach series 173 (DE-604)BV007921321 173 HEBIS Datenaustausch Darmstadt application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=007582043&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Subcellular fractionation a practical approach The practical approach series Cellen (biologie) gtt Fractionering gtt Weefsel gtt Subcellular Fractions analysis Subcellular Fractions methods Subcellular fractionation Handbooks, manuals, etc Subcellular fractionation Laboratory manuals Zelle (DE-588)4067537-3 gnd Kompartiment (DE-588)4426443-4 gnd Fraktionierung (DE-588)4018056-6 gnd Methode (DE-588)4038971-6 gnd |
| subject_GND | (DE-588)4067537-3 (DE-588)4426443-4 (DE-588)4018056-6 (DE-588)4038971-6 |
| title | Subcellular fractionation a practical approach |
| title_auth | Subcellular fractionation a practical approach |
| title_exact_search | Subcellular fractionation a practical approach |
| title_full | Subcellular fractionation a practical approach ed. by J. M. Graham ... |
| title_fullStr | Subcellular fractionation a practical approach ed. by J. M. Graham ... |
| title_full_unstemmed | Subcellular fractionation a practical approach ed. by J. M. Graham ... |
| title_short | Subcellular fractionation |
| title_sort | subcellular fractionation a practical approach |
| title_sub | a practical approach |
| topic | Cellen (biologie) gtt Fractionering gtt Weefsel gtt Subcellular Fractions analysis Subcellular Fractions methods Subcellular fractionation Handbooks, manuals, etc Subcellular fractionation Laboratory manuals Zelle (DE-588)4067537-3 gnd Kompartiment (DE-588)4426443-4 gnd Fraktionierung (DE-588)4018056-6 gnd Methode (DE-588)4038971-6 gnd |
| topic_facet | Cellen (biologie) Fractionering Weefsel Subcellular Fractions analysis Subcellular Fractions methods Subcellular fractionation Handbooks, manuals, etc Subcellular fractionation Laboratory manuals Zelle Kompartiment Fraktionierung Methode |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=007582043&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| volume_link | (DE-604)BV007921321 |
| work_keys_str_mv | AT grahamjohnm subcellularfractionationapracticalapproach |