Chromatin and Chromosomal Protein Research II

Name: Chromatin and Chromosomal Protein Research II
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Academic Press

Published on 26. July 1978

433 pages

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978-0-08-085914-9 (ISBN)

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Front Cover
Methods in Cell Biology, Volume XVII
Copyright Page
Contents
List of Contributers
Preface
Erratum
Part A: Isolation of Nuclei and Preparation of Chromatin. II
Chapter I. Procedures for Minimizing Protease Activity during Isolation of Nuclei, Chromatin, and the Histones
I. Introduction
II. Methods
III. Conditions and Patterns of Proteolytic Degradation of Histones
IV. Concluding Remarks
References
Chapter 2. Isolation of Nuclei from Animal Cells in Culture
I. Introduction
II. General Consideration of Nuclear Isolation Procedures
III. Methods for Isolating Nuclei
References
Chapter 3. Preparation of Chromatin from Animal Tissues and Cultured Cells
I. Introduction
II. Rationales of Methods
III. Technical Considerations
IV. Direct Methods of Chromatin Isolation
V. Chromatin Isolation from Nuclei
VI. Fragmentation of Chromatin
VII. Storage
VIII. Criteria of Purity
IX. Criteria of Structural Preservation
X. Choice of Methods
Note Added in Proof
References
Chapter 4. Methods for Isolation of Nuclei from Cultured Mammalian Cells
Conditions for Preferential Retention of Selected Histones
I. Introduction
II. Method A: For Studies on Lysine-Rich and Slightly Lysine-Rich Histones
III. Method B: For Studies on Arginine-Rich Histones
References
Chapter 5. Methods for Isolation of Nuclei from Spermatozoa
I. Introduction
II. Spermatozoa of Eutherian Mammals
III. Spermatozoa with Protamine
IV. Spermatozoa with Somatic Histone-Type Protein
V. Spermatozoa of Molluscs
VI. Spermatozoa of Cricket
VII. Spermatozoa of Crab
References
Chapter 6. Manual Enucleation of Xenopus Oocytes
I. Introduction
II. Methods
III. Conclusions
References
Chapter 7. Macro and Micro Methods for Isolation and Injection of Cell Components from Polytene Tissues
I. Mass Isolation of Salivary Glands from Drosophila hydei
II. Micromanipulation of Salivary Glands
III. Microinjection of Salivary Gland Cells
References
Chapter 8. Isolation of Nuclei and Chromatin from Phycomyces blakesleeanus
I. Introduction
II. Methods
References
Part B: Chromosomes
Chapter 9. Mammalian Metaphase Chromosomes
I. Introduction
II. General Procedures
III. Isolation of Chromosomes
IV. Chromosome Aggregation
References
Chapter 10. Analytical Techniques for Isolated Metaphase Chromosome Fractions
I. Introduction
II. Chromosome Isolation
III. Fractionation of Isolated Chromosomes
IV. Fraction Analysis
V. Computer Analysis
References
Chapter 11. Methods and Mechanisms of Chromosome Banding
I. Introduction
II. Slide Preparation
III. Q-Banding
IV. G-Banding
V. C-Banding
VI. R-Banding
VII. BG-Banding
References
Chapter 12. Cytological Analysis of Drosophila Polytene Chromosomes
I. Introduction
II. Staining with Aceto-orcein and Fast Green FCF
III. Staining with Naphtol-Yellow S (NYS)
IV. The Squash and Selection Technique
References
Chapter 13. Isolation and Purification of Nucleoli and Nucleolar Chromatin from Mammalian Cells
I. Introduction
II. Isolation of Nucleoli from Various Mammalian Cells
III. Purification of Nucleolar Chromatin
IV. Commentary
References
Chapter 14. Nucleolar Proteins
I. Introduction
II. Direct Extraction of Nucleolar Proteins
III. Proteins of Nucleolar Subcomponents
IV. Analyses and Fractionation of Nucleolar Proteins
V. Nucleolar Enzymes
VI . Methods for Studies on Nucleolar Antigens
VII. Discussion
References
Part C: Fractionation and Characterization of Histones. II
Chapter 15. Electrophoretic Fractionation of Histones Utilizing Starch Gels and Sodium Dodecyl Sulfate-Urea Gels
I. Introduction
II. Starch Gel Electrophoresis
III. Histone Electrophoresis in Polyacrylamide Gels Containing SDS
References
Chapter 16. Resolution of Histones by Polyacrylamide Gel Electrophoresis in Presence of Nonionic Detergents
I. Introduction
II. Method
III. Interpretation and Evaluation
IV. Applications
References
Chapter 17. Polyacrylamide Gel Electrophoretic Fractionation of Histones
I. Introduction
II. Low pH Gel Systems
III. High pH-SDS Gel Systems
IV. Neutral pH Systems
V. Two-Dimensional Gel Systems
VI. Concluding Remarks
References
Part D: Fractionation and Characterization of Nonhistone Chromosomal Proteins
Chapter 18. Affinity Chromatography of DNA-Binding Proteins on DNA Covalently Attached to Solid Supports
I. Introduction
II. Preparation of DNA Columns
III. Covalent Coupling of DNA to Polysaccharide Supports
IV. Deproteinization of DNA Samples
V. DNA Shearing, Sizing, and Cot Fractionation
VI. Coupling Reaction for Double-Stranded DNA
VII. Coupling Reaction for Single-Stranded DNA
VIII. Properties of DNA-Affinity Columns
IX. DNA-Affinity Chromatography of Nuclear Proteins
X. Concentration Dependence and Saturability of Protein-DNA Binding
XI. Ionic-Strength Dependence of Protein-DNA Binding
XII. DNA Binding by Nuclear Proteins Extractable in 0.35 M NaCI-A Basis for Fractionation
XIII. Evidence for Sequence Specificity in DNA Binding by Nuclear Nonhistone Proteins
References
Chapter 19. DNA-Binding Proteins
I. Introduction
II. DNA-Cellulose Chromatography
III. DNA-Agarose and DNA-Acrylamide Chromatography
IV. Discussion
References
Chapter 20. Fractionation of Nonhistone Proteins by Histone-Affinity Chromatography
I. Introduction
II. Methods
III. Results and Discussion
References
Chapter 21. Fractionation of Nonhistone Chromosomal Proteins
I. Introduction
II. General Considerations
III. Selective Extraction of Nonhistone Chromosomal Proteins
IV. Nonhistone Chromosomal Protein Fractionation Procedures
V. QAE-Sephadex Fractionation of Nonhistone Chromosomal Proteins
VI. Assessment of Protein Purification
References
Chapter 22. Fractionation of Chromosomal Nonhistone Proteins Using Chromatin-Cellulose Resins: Purification of Steroid Hormone Acceptor Proteins
I. Introduction
II. Materials
III. Method for Preparation of Chromatin-Cellulose, NAP-Cellulose, and DNA-Cellulose Resins
IV. Analysis of Resins
V. Selection of Conditions for Maximal Binding of Chromatin, NAP (or Pure DNA) to Cellulose
VI. Integrity of Chromatin and Nucleoproteins Bound to Cellulose
VII. Fractionation of Chromosomal Protein from Chromatin-Cellulose Resins
VIII. Isolation of the Nuclear Acceptors Which Bind Steroid Hormones
IX. Comments on the Technique
References
Chapter 23. Methods for Assessing the Binding of Steroid Hormones in Nuclei
I. Introduction
II. Receptor Binding in Isolated Nuclei
III. Nuclear Exchange Assays
Note Added in Proof
References
Chapter 24. Methods for Assessing the Binding of Steroid Hormones in Nuclei and Chromatin
I. Introduction
II. Steroid Exchange Methodology
References
Chapter 25. Proteins of Nuclear Ribonucleoprotein Subcomplexes
I. Introduction
II. Expectations, Methods, and Criteria
III. Protein Composition of hnRNP
IV. Nuclear Poly(A) Containing Ribonucleoproteins
V. Association of Ribonucleoprotein with Chromatin
References
Chapter 26. Isolation and Characterization of Ribonucleoprotein Particles Containing Heterogeneous Nuclear RNA
I. Introduction
II. Isolation of Nuclear hn RNP Particles
III. Physiochemical Characterization of HeLa hn RNP Particles
IV. Sources of Chemicals
References
Chapter 27. Chromosomal Association of an Epstein-Barr Virus-Associated Nuclear Antigen
I. Introduction
II. Methods for Detection of EBNA
III. Visualization of EBNA Metaphase Chromosomes
IV. Discussion on Methods
V. EBNA Is a Chromatin-Associated Protein
References
Subject Index
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Chromatin and Chromosomal Protein Research II

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