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Book Cover
E-book
Author Graham, J. M

Title Subcellular Fractionation : A Practical Approach
Published Oxford : Oxford University Press, 1997
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Description 1 online resource (360 p.)
Series Practical Approach Series ; v.173
Practical approach series
Contents Cover -- Contents -- List of contributors -- Abbreviations -- 1. Homogenization of tissues and cells -- 1. Introduction -- 2. Aims of the homogenization procedure -- 3. Influence of sample type -- 4. Homogenization media -- 5. Methods of homogenization -- Type 1 homogenizers -- Type 2 homogenizers -- 6. Homogenization of tissues and cells -- Mammalian liver -- Brain -- Muscle -- Mammalian tissue culture cells -- Plant organelles -- Yeast -- Other fungi and algae -- Trypanosomes -- Bacteria -- References -- 2. Isolation of subcellular fractions -- 1. Introduction
1. Introduction -- 2. Methods of preparing purified nuclei -- Types of cells and tissue samples -- Homogenization media -- Homogenization methods -- Centrifugation conditions -- Assays of nuclear purity -- 3. Methods for purifying metaphase chromosomes -- 4. Isolation of nuclear subfractions -- Preparation of nucleoli -- Preparation of nuclear membranes -- Isolation of nuclear matrix -- Preparation of nucleoids -- 5. Isolation of nucleoprotein complexes -- Isolation of polynucleosomes of chromatin -- Ribonucleoproteins -- 6. Isolation of nuclear macromolecules -- Isolation of nuclear proteins
2. Composition of a tissue homogenate -- 3. Properties of cell organelles -- Factors affecting organelle density and size -- 4. Centrifugal methods for the separation of organelles -- Separation by size -- Separation by density -- Density perturbation -- 5. Non-centrifugal procedures -- Immunoisolation -- Separation by electrophoresis -- 6. Identification of separated material -- Marker enzymes -- Introduced markers for endocytic and exocytic pathways -- Characteristic non-enzymatic proteins -- 7. Assessment of the purity of fractions -- Purity and purification
Determination of the integrity of the mitochondrial outer membrane
Isolation of nuclear RNA -- Isolation of DNA -- 7. Functional assays of nuclei -- Analysis of DNA-binding proteins -- Transcription assays -- References -- 4. Subcellular fractionation of mitochondria -- 1. Introduction -- 2. Purification of mitochondria from various eukaryotic sources -- Introduction -- Protocols for purification of mitochondria from several eukaryotic sources -- Further purification of mitochondrial fractions -- 3. Determination of mitochondrial purity -- Introduction -- Use of the oxygen electrode to determine mitochondrial integrity
Problems from cell heterogeneity within tissues -- Problems arising from organelle fragmentation -- Missorting in the exocytic and endocytic pathways -- 8. Fractionation problems -- No separation -- Aggregation following resuspension of a fraction -- Poor recovery of markers -- Damage to cell structures -- 9. A systematic approach to cell fractionation -- Preliminary studies -- Determination of the properties of components of the homogenate -- Method development -- Simplification of the separation -- References -- 3. Isolation and characterization of nuclei and nuclear subfractions
Summary Many investigations into the structure and function of cells and tissues require the isolation of a particular membrane or subcellular component (organelle). This book covers all the necessary aspects, from breaking up the cells (homogenization), via a variety of separation techniques (the isolation and fractionation chapters), to characterization of the separated organelles
Notes Description based upon print version of record
ProQuest Ebook Central Rental
Form Electronic book
Author Rickwood, D. (David)
ISBN 9780191591617
OTHER TI ProQuest Ebook Central