Chapter 4. Isolation and Functional Analysis of Organelles
- J. Robin Harris1,
- John Graham2,
- David Rickwood3
Published Online: 3 JUL 2006
DOI: 10.1002/0470033487.ch4
Copyright © 2006 John Wiley & Sons, Ltd
Book Title
Cell Biology Protocols
Additional Information
How to Cite
Graham, J. and Harris, J. R. (2006) Isolation and Functional Analysis of Organelles, in Cell Biology Protocols (eds J. R. Harris, J. Graham and D. Rickwood), John Wiley & Sons, Ltd, Chichester, UK. doi: 10.1002/0470033487.ch4
Editor Information
- 1
Institute of Zoology, Johannes Gutenberg-Universität, University of Mainz, D-55099 Mainz, Germany
- 2
JG Research Consultancy, 34 Meadway, Upton Wirral CH49 6IQ, UK
- 3
Department of Biological Sciences, University of Essex, Wivenhoe Park, Colchester CO4 3SQ, UK
Publication History
- Published Online: 3 JUL 2006
- Published Print: 27 JAN 2006
ISBN Information
Print ISBN: 9780470847589
Online ISBN: 9780470033487
- Summary
- Chapter
Keywords:
- differential centrifugation;
- density gradient centrifugation;
- enzyme marker;
- chloroplast functional assays;
- nuclear pore complex isolation;
- yeast mitochondria purification;
- catalase assay;
- human erythrocyte ghosts;
- assessing chloroplast integrity
Summary
This chapter contains sections titled:
Introduction
Homogenization
Differential centrifugation
Density gradient centrifugation
Nuclei and nuclear components
Mitochondria
Lysosomes
Peroxisomes
Rough and smooth endoplasmic reticulum (ER)
Golgi membranes
Plasma membrane
Chloroplasts
Protocols 4.1 Isolation of nuclei from mammalian liver in an iodixanol gradient (with notes on cultured cells)
Protocols 4.2 Isolation of metaphase chromosomes
Protocols 4.3 Isolation of the nuclear envelope
Protocols 4.4 Nuclear pore complex isolation
Protocols 4.5 Preparation of nuclear matrix
Protocols 4.6 Preparation of nucleoli
Protocols 4.7 Isolation of a heavy mitochondrial fraction from rat liver by differential centrifugation
Protocols 4.8 Preparation of a light mitochondrial fraction from tissues and cultured cells
Protocols 4.9 Purification of yeast mitochondria in a discontinuous Nycodenz® gradient
Protocols 4.10 Purification of mitochondria from mammalian liver or cultured cells in a median-loaded discontinuous Nycodenz® gradient
Protocols 4.11 Succinate–INT reductase assay
Protocols 4.12 Isolation of lysosomes in a discontinuous Nycodenz® gradient
Protocols 4.13 β-Galactosidase (spectrophotometric assay)
Protocols 4.14 β-Galactosidase (fluorometric assay)
Protocols 4.15 Isolation of mammalian peroxisomes in an iodixanol gradient
Protocols 4.16 Catalase assay
Protocols 4.17 Analysis of major organelles in a preformed iodixanol gradient
Protocols 4.18 Separation of smooth and rough ER in preformed sucrose gradients
Protocols 4.19 Separation of smooth and rough ER in a self-generated iodixanol gradient
Protocols 4.20 NADPH-cytochrome c reductase assay
Protocols 4.21 Glucose-6-phosphatase assay
Protocols 4.22 RNA analysis
Protocols 4.23 Isolation of Golgi membranes from liver
Protocols 4.24 Assay of UDP-galactose galactosyl transferase
Protocols 4.25 Purification of human erythrocyte ‘ghosts’
Protocols 4.26 Isolation of plasma membrane sheets from rat liver
Protocols 4.27 Assay for 5'-nucleotidase
Protocols 4.28 Assay for alkaline phosphodiesterase
Protocols 4.29 Assay for ouabain-sensitive Na+/K+-ATPase
Protocols 4.30 Isolation of chloroplasts from green leaves or pea seedlings
Protocols 4.31 Measurement of chloroplast chlorophyll
Protocols 4.32 Assessment of chloroplast integrity
References