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In Situ Detection of DNA Damage: Methods and Protocols: Methods in Molecular Biology, cartea 203

Editat de Vladimir V. Didenko
en Limba Engleză Paperback – 10 noi 2010
Detection and analysis of DNA damage is of critical importance in a variety of biological disciplines studying apoptosis, cell cycle and cell di- sion, carcinogenesis, tumor growth, embryogenesis and aging, neu- degenerative and heart diseases, anticancer drug development, environmental and radiobiological research, and others. Individual cells within the same tissue or in cell culture may vary in the extent of their DNA damage and, consequently, can display different re- tions to it. These differences between individual cells in the same cell popu- tion are detected using in situ approaches. In situ is a Latin term meaning “on site” or “in place.” It is used to denote the processes occurring or detected in their place of origin. In mole- lar and cell biology this usually refers to undisrupted mounted cells or tissue sections. In that meaning “in situ” is used as part of the terms “in situ PCR,” “in situ transcription,” “in situ hybridization,” “in situ end labeling,” and “in situ ligation.” Sometimes the “in situ” term is applied at the subcellular level to cells disrupted in the process of analysis, for example, in the detection of specific sequences in chromosomes using fluorescent in situ hybridization (FISH). Historically, the term was used primarily in methods dealing with nucleic acids.
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Specificații

ISBN-13: 9781617372728
ISBN-10: 1617372722
Pagini: 336
Ilustrații: XVII, 313 p. 164 illus., 31 illus. in color.
Dimensiuni: 152 x 229 x 18 mm
Greutate: 0.45 kg
Ediția:Softcover reprint of hardcover 1st ed. 2002
Editura: Humana Press Inc.
Colecția Humana
Seria Methods in Molecular Biology

Locul publicării:Totowa, NJ, United States

Public țintă

Professional/practitioner

Cuprins

Labeling DNA Breaks Using Terminal Transferase (TUNEL Assay).- Labeling DNA Damage with Terminal Transferase.- TUNEL Assay.- Electron Microscopic Detection of DNA Damage Labeled by TUNEL.- Quantitative Differentiation of Both Free 3? OH and 5? OH DNA Ends Using Terminal Transferase-Based Labeling Combined with Transmission Electron Microscopy.- Determination of Three-Dimensional Distribution of Apoptotic DNA Damage by Combination of TUNEL and Quick-Freezing and Deep-Etching Techniques.- In Situ Detection of DNA Strand Breaks in Analysis of Apoptosis by Flow- and Laser-Scanning Cytometry.- Labeling DNA Breaks Using DNA Polymerase I or its Klenow Fragment.- DNA Damage Detection Using DNA Polymerase I or its Klenow Fragment.- Labeling DNA Breaks In Situ by Klenow Enzyme.- In Situ Nick Translation at the Electron Microscopic Level.- Labeling DNA Breaks Using Ligase.- In Situ DNA Ligation as a Method for Labeling Apoptotic Cells in Tissue Sections.- Detection of Specific Double-Strand DNA Breaks and Apoptosis In Situ Using T4 DNA Ligase.- In Situ Detection of Double-Strand DNA Breaks with Terminal 5?OH Groups.- Detection of DNA Breaks in Agarose Trapped Cells: Comet Assay and Related Techniques.- The Comet Assay.- The Comet Assay.- Ultrasensitive Detection of DNA Damage by the Combination of the Comet and TUNEL Assays.- Application of FISH to Detect DNA Damage.- Detection of Modified Bases and AP Sites in DNA.- Simultaneous In Situ Detection of DNA Fragmentation and RNA/DNA Oxidative Damage Using TUNEL Assay and Immunohistochemical Labeling for 8-Hydroxy-2?-Deoxyguanosine (8-OHdG).- The In Situ Detection of Apurinic/Apyrimidinic Sites and DNA Breaks Bearing Extension Blocking Termini.- Indirect and General Markers of DNA Damage.- Markers of Poly (ADP-Ribose)Polymerase Activity as Correlates of DNA Damage.- Ultrasound Imaging of Apoptosis.- p53 Induction as an Indicator of DNA Damage.- Detection of Caspases Activation In Situ by Fluorochrome-Labeled Inhibitors of Caspases (FLICA).

Recenzii

"Investigators working in the general area of DNA damage, including apoptosis, will find this book of use. The protocols are sufficiently detailed so as to allow experienced laboratory workers access to the described methods. . .the broad spectrum of methods described allows for individual investigators to exercise options."-Doody's Health Sciences Book Review Journal

"Here, a panel of experts describe all the major in situ techniques for studying DNA damage and apoptosis, and how approaches originally designed to label apoptotic cells can be used for DNA damage analysis (and vice versa)." - European Journal of Histochemistry

Textul de pe ultima copertă

Detection and analysis of DNA damage is critically important in the study of the widest variety of biological phenomena, ranging from apoptosis and aging to heart diseases and anticancer drug development. In In Situ Detection of DNA Damage: Methods and Protocols, Vladimir Didenko and a panel of experts describe all the major in situ techniques for studying DNA damage and apoptosis, and show how approaches originally designed to label apoptotic cells can be used for DNA damage analysis (and vice versa). The readily reproducible techniques presented here include all those designed to detect specific single- and double-stranded DNA breaks in tissue sections while using polymerases, ligases, nucleases, and kinases. In-depth protocols for single-cell electrophoresis (comet assay) are presented, as well as those for the detection of indirect markers of DNA damage and modified bases. Technical reviews discuss specificity, sensitivity, advantages, and limitations of the techniques described, and compare them with alternative approaches.
State-of-the-art and highly practical, In Situ Detection of DNA Damage: Methods and Protocols presents for the first time a complete set of enzymatic methods for determining DNA breaks, and will greatly facilitate the work of all those studying not only the in situ detection of DNA damage but also the labeling of apoptotic cells.

Caracteristici

Includes supplementary material: sn.pub/extras