Nuclear structures: isolation and characterization
Gespeichert in:
| Format: | Buch |
|---|---|
| Sprache: | English |
| Veröffentlicht: |
London u.a.
Butterworths
1986
|
| Ausgabe: | 1. publ. |
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | 209 S. Ill. |
| ISBN: | 0407003231 |
Internformat
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| 245 | 1 | 0 | |a Nuclear structures |b isolation and characterization |c ed. by A. J. MacGillivray ... |
| 250 | |a 1. publ. | ||
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| 650 | 4 | |a Cell fractionation | |
| 650 | 4 | |a Cell nuclei |x Separation | |
| 650 | 4 | |a Cell organelles | |
| 650 | 4 | |a Cytochemistry | |
| 650 | 4 | |a Subcellular Fractions |x chemistry | |
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Datensatz im Suchindex
| _version_ | 1804114965194342400 |
|---|---|
| adam_text | Nuclear structures
Isolation and characterization
Edited by
A J MacGillivray
Reader in Biochemistry
School of Biological Sciences, University of Sussex
and
G D Birnie
Senior Scientist
Beatson Institute for Cancer Research, Glasgow
Butterworths
London • Boston • Durban • Singapore • Sydney • Toronto • Wellington
Contents
Preface v
Contributors vii
1 Nuclear matrices 1
ShonaA Comerford, PaulS Agutter and Alexander G McLennan
Isolation of nuclear matrices 2
Conclusions 9
Notes on the methods 9
Acknowledgements 12
References 12
2 Nucleoids 14
Dean A Jackson
Nucleoid preparation and morphology 14
Preparation of nucleoids from HeLa cells 14
HeLa nucleoid morphology 15
Preparation of nucleoids from other cell types 18
Procedures for stabilizing nucleoids 23
Characterization of nucleoids 23
Proteins 23
Enzyme activities 24
DNA 26
RNA 28
Experimental uses of nucleoids 29
Criticism of nucleoid preparation 30
Conclusions 31
Acknowledgement 32
References 32
3 Nuclear envelopes 34
PaulS Agutter
Background 34
Objectives of nuclear envelope isolation 34
General constraints 35
Development of isolation procedures to meet different research
objectives 36
Procedures 36
High ionic strength procedures 36
Low ionic strength procedures 39
General considerations 41
Notes on the methods 43
References 45
4 Polynucleosomes and monomer nucleosomes 47
/ Allan, R H Nicolas and G H Goodwin
Isolation of polynucleosomes 48
General points on the preparation of polynucleosomes 49
The preparation of chicken erythrocyte polynucleosomes 56
Fractionation of monomer nucleosomes 59
Isolation of salt-soluble nucleosomes 60
Electrophoresis of nucleosomes 61
Analysis of electrophoretically separated nucleosomes 63
Evaluation of methods used to isolate transcriptionally active
nucleosomes 68
Acknowledgements 71
References 71
5 Metaphase chromosomes 74
Bryan D Young
Bulk preparation of metaphase chromosomes 75
Details of procedures 75
Discussion of methods 77
Flow cytometry of metaphase chromosomes 79
Choice of DNA-specific stain 79
Technical innovations 80
Applications of chromosome fractionation 81
Flow karyotype analysis 81
Gene mapping 82
Chromosomal DNA library construction 83
Acknowledgements 84
References 84
6 DNA tight-binding proteins 86
Ailsa M Campbell and Caroline V P Addey
The definition of a DNA tight-binding protein 86
Definition based on analogy with bacterial systems 86
Operational definition based on isolation procedures 86
Operational definition based on specific DNA binding 87
The preparation of DNA tight-binding proteins 87
Isolation of nuclei 87
Isolation of chromatin 89
Extraction of DNA tight-binding proteins 89
Immunological characterization of tight-binding proteins 92
Methods of immunoassay 92
DNA binding characteristics 95
Detection of sequence specific DNA binding proteins 96
Conclusions 98
Acknowledgements 98
References 98
7 Nucleoli and preribosomal ribonucleoprotein particles 100
Trevor J C Beebee
Methods for the isolation of nucleoli 100
Isolation using ultrasonication 100
Isolation using homogenization 102
Other methods 104
Assessment of nucleolar preparations 106
Yield and purity 106
Internal integrity 106
Conclusions 111
Isolation of preribosomal ribonucleoprotein particles 112
Methods 112
Assessment of preparations 113
Overview 116
Acknowledgements 116
References 116
8 Ribonucleoprotein particles containing heterogeneous nuclear RNA 118
J T Knowler, C W McGregor and Z Islam
Isolation of hnRNP 118
Isolation of hnRNP by extraction from nuclei 118
Isolation of hnRNP by nuclear lysis 120
Isolation of monomeric versus polymeric hnRNP 122
Characterization of hnRNP 122
The purity and authenticity of isolated hnRNP 122
Further purification of hnRNP: the problem of desirability 124
The fractionation and characterization of hnRNP proteins 124
Acknowledgement 128
References 128
9 Nuclear RNP particles containing small RNAs 130
Alexander J MacGillivray
Background, objectives and limitations to the preparation of snRNP
particles 131
Procedures using immunoaffinity chromatography 135
Preparation of snRNP particles containing Ul and U2 4-6 snRNAs 135
Preparation of La-RNP 139
Immunoaffinity chromatography using autoantibodies - conclusions 142
Use of antibodies to 2,2,7-trimethylguanosine to isolate snRNP
particles 143
Procedures using non-immunological techniques 148
Isolation of total snRNP particles and their partial fractionation 148
Preparation of Ul, Ul-2 and U4-6 snRNP particles 150
Preparation of Ul and U2 snRNP particles 154
General conclusions 157
Acknowledgements 160
References 160
10 Hormone receptors 163
Robin Leake
Peptide hormones 163
Thyroid hormones 164
Steroid hormones 164
The classic model 165
The equilibrium model 166
A new model of steroid receptor action 169
Properties of receptors for individual steroids 171
Assay methods 175
Acknowledgements 178
References 179
11 DNA and RNA 182
G D Birnie and S V Graham
Isolation of nuclei 183
Need for isolated nuclei 183
Rationales of protocols 184
Fractionation of cells 185
Preparation of purified nuclei 187
Isolation of nucleic acids 189
DNA 189
RNA 191
Polyadenylated RNA 194
Assessment of preparations 196
Nuclei 196
DNA 197
RNA 198
Acknowledgements 200
References 200
Index 203
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| indexdate | 2024-07-09T15:15:06Z |
| institution | BVB |
| isbn | 0407003231 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-000318115 |
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| physical | 209 S. Ill. |
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| spelling | Nuclear structures isolation and characterization ed. by A. J. MacGillivray ... 1. publ. London u.a. Butterworths 1986 209 S. Ill. txt rdacontent n rdamedia nc rdacarrier Nucleo (Citologia) larpcal Cell Fractionation methods Cell Nucleus ultrastructure Cell fractionation Cell nuclei Separation Cell organelles Cytochemistry Subcellular Fractions chemistry Struktur (DE-588)4058125-1 gnd rswk-swf Zellkern (DE-588)4067544-0 gnd rswk-swf (DE-588)4143413-4 Aufsatzsammlung gnd-content Zellkern (DE-588)4067544-0 s Struktur (DE-588)4058125-1 s DE-604 MacGillivray, Alexander J. Sonstige oth HEBIS Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=000318115&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Nuclear structures isolation and characterization Nucleo (Citologia) larpcal Cell Fractionation methods Cell Nucleus ultrastructure Cell fractionation Cell nuclei Separation Cell organelles Cytochemistry Subcellular Fractions chemistry Struktur (DE-588)4058125-1 gnd Zellkern (DE-588)4067544-0 gnd |
| subject_GND | (DE-588)4058125-1 (DE-588)4067544-0 (DE-588)4143413-4 |
| title | Nuclear structures isolation and characterization |
| title_auth | Nuclear structures isolation and characterization |
| title_exact_search | Nuclear structures isolation and characterization |
| title_full | Nuclear structures isolation and characterization ed. by A. J. MacGillivray ... |
| title_fullStr | Nuclear structures isolation and characterization ed. by A. J. MacGillivray ... |
| title_full_unstemmed | Nuclear structures isolation and characterization ed. by A. J. MacGillivray ... |
| title_short | Nuclear structures |
| title_sort | nuclear structures isolation and characterization |
| title_sub | isolation and characterization |
| topic | Nucleo (Citologia) larpcal Cell Fractionation methods Cell Nucleus ultrastructure Cell fractionation Cell nuclei Separation Cell organelles Cytochemistry Subcellular Fractions chemistry Struktur (DE-588)4058125-1 gnd Zellkern (DE-588)4067544-0 gnd |
| topic_facet | Nucleo (Citologia) Cell Fractionation methods Cell Nucleus ultrastructure Cell fractionation Cell nuclei Separation Cell organelles Cytochemistry Subcellular Fractions chemistry Struktur Zellkern Aufsatzsammlung |
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