Cell cycle control and dysregulation protocols : cyclins, cyclin-dependent kinases, and other factors / edited by Antonio Giordano, Gaetano Romano.

Contributor(s): Giordano, Antonio, MD | Romano, Gaetano, 1959-
Material type: TextTextSeries: Methods in molecular biology (Clifton, N.J.): v. 285.Publisher: Totowa, N.J. : Humana Press, ©2004Description: 1 online resource (xiii, 185 pages) : illustrations (some color)Content type: text Media type: computer Carrier type: online resourceISBN: 0896039498; 9780896039490; 1592598226; 9781592598229Subject(s): Cell cycle -- Laboratory manuals | Cyclin-dependent kinases -- Laboratory manuals | Cyclins -- Laboratory manuals | Biology | Immunohistochemistry | Nucleic Acids | Protein-Serine-Threonine Kinases | Genetic Phenomena | Proline-Directed Protein Kinases | Proteins | Peptides | Immunologic Tests | Amino Acids, Peptides, and Proteins | Biological Science Disciplines | Protein Kinases | Histocytochemistry | Nucleic Acids, Nucleotides, and Nucleosides | Chemicals and Drugs | Natural Science Disciplines | Immunologic Techniques | Phenomena and Processes | Histological Techniques | Laboratory Techniques and Procedures | Phosphotransferases (Alcohol Group Acceptor) | Diagnosis | Clinical Laboratory Techniques | Phosphotransferases | Investigative Techniques | Disciplines and Occupations | Analytical, Diagnostic and Therapeutic Techniques and Equipment | Transferases | Enzymes | Enzymes and Coenzymes | Cell Cycle Proteins | DNA | Gene Expression | Fluorescent Antibody Technique | Cyclin-Dependent Kinases | Intracellular Signaling Peptides and Proteins | Genetics | SCIENCE -- Life Sciences -- Developmental Biology | Cell cycle | Cyclin-dependent kinases | CyclinsGenre/Form: Electronic books. | Laboratory manuals. Additional physical formats: Print version:: Cell cycle control and dysregulation protocols.DDC classification: 571.8/4 LOC classification: QH605 | .C4257 2004Online resources: Click here to access online
Contents:
The biology of cyclins and cyclin-dependent protein kinases: an introduction / Lucio Miele -- In situ immunofluorescence analysis: immunofluorescence microscopy / Amjad Javed [and others] -- In situ immunofluorescence analysis: analyzing RNA synthesis by 5-bromouridine-5'-triphosphate labeling / Amjad Javed [and others] -- Immunofluorescence analysis using epitope-tagged proteins: in vitro system / Amjad Javed [and others] -- Analysis of in vivo gene expression using epitope-tagged proteins / Amjad Javed [and others] -- Chromatin immunoprecipitation / Amjad Javed [and others].
Protein/deoxyribonucleic acid interactions linked to gene expression: electrophoretic mobility shift assay / Amjad Javed [and others] -- Protein/deoxyribonucleic acid interactions linked to gene expression: DNase I digestion / Amjad Javed [and others] -- Protein/deoxyribonucleic acid interactions linked to gene expression: ligation-mediated polymerase chain reaction / Amjad Javed [and others] -- Assays for cyclin-dependent kinase inhibitors / Adrian M. Senderowicz and Tyler Lahusen -- Protein degradation via the proteosome / Henry Hoff, Hong Zhang, and Christian Sell.
The transformed phenotype / Henry Hoff [and others] -- A morphologic approach to detect apoptosis based on electron microscopy / Martyn K. White, Timothy G. Schneider, and Caterina Cinti -- Detection of apoptotic deoxyribonucleic acid break by in situ / Nick Translation [and others] -- Induction of deoxyribonucleic acid damage by alkylating agents / Salvatore Cortellino [and others] -- Induction of deoxyribonucleic acid damage by -- irradiation / Salvatore Cortellino, David P. Turner and Alfonso Bellacosa -- Ultraviolet irradiation of cells / David P. Turner, Anthony T. Yeung, and Alfonso Bellacosa -- Transient production of retroviral- and lentiviral-based vectors for the transduction of mammalian cells / Tiziana Tonini [and others].
Determination of functional viral titer by drug-resistance -- Colony assay, expression of green fluorescent protein, and '-galactoside staining / Tiziana Tonini [and others] -- Retroviral and lentiviral vector titration by the analysis of the activity of viral reverse transcriptase / Tiziana Tonini [and others] -- Single and double colloidal gold labeling in postembedding immunoelectron microscopy / Nicoletta Zini [and others] -- Multifluorescence labeling and colocalization analyses / Massimo Riccio [and others].
Summary: The dysregulation of the normal cell cycle underlies an enormous number of pathological conditions, including the neurodegenerative disorders, all cancers, and diabetes. In Cell Cycle Control and Dysregulation Protocols: Cyclins, Cyclin-Dependent Kinases, and Other Factors, expert laboratorians detail emerging methodologies for studying the cell cycle, its kinases, and kinase inhibitors. The authors focus on issues of gene expression in vivo and in vitro, the analysis of cyclin-dependent kinase inhibitors, protein degradation mediated by the proteosome, the analysis of the transformed cell phenotype, and innovative techniques to detect apoptosis. Many of the protocols are new, such as electron microsocopy to detect apoptosis and proteosome-mediated degradation, whereas others study the interactions between protein-protein, protein-DNA, and protein-RNA. The protocols follow the successful Methods in Molecular Biology™ series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. State-of-the-art and highly practical, Cell Cycle Control and Dysregulation Protocols: Cyclins, Cyclin-Dependent Kinases, and Other Factors offers pathbreaking researchers powerful tools for probing the cell cycle regulation across a wide variety of pathologies.
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Includes bibliographical references and index.

The biology of cyclins and cyclin-dependent protein kinases: an introduction / Lucio Miele -- In situ immunofluorescence analysis: immunofluorescence microscopy / Amjad Javed [and others] -- In situ immunofluorescence analysis: analyzing RNA synthesis by 5-bromouridine-5'-triphosphate labeling / Amjad Javed [and others] -- Immunofluorescence analysis using epitope-tagged proteins: in vitro system / Amjad Javed [and others] -- Analysis of in vivo gene expression using epitope-tagged proteins / Amjad Javed [and others] -- Chromatin immunoprecipitation / Amjad Javed [and others].

Protein/deoxyribonucleic acid interactions linked to gene expression: electrophoretic mobility shift assay / Amjad Javed [and others] -- Protein/deoxyribonucleic acid interactions linked to gene expression: DNase I digestion / Amjad Javed [and others] -- Protein/deoxyribonucleic acid interactions linked to gene expression: ligation-mediated polymerase chain reaction / Amjad Javed [and others] -- Assays for cyclin-dependent kinase inhibitors / Adrian M. Senderowicz and Tyler Lahusen -- Protein degradation via the proteosome / Henry Hoff, Hong Zhang, and Christian Sell.

The transformed phenotype / Henry Hoff [and others] -- A morphologic approach to detect apoptosis based on electron microscopy / Martyn K. White, Timothy G. Schneider, and Caterina Cinti -- Detection of apoptotic deoxyribonucleic acid break by in situ / Nick Translation [and others] -- Induction of deoxyribonucleic acid damage by alkylating agents / Salvatore Cortellino [and others] -- Induction of deoxyribonucleic acid damage by -- irradiation / Salvatore Cortellino, David P. Turner and Alfonso Bellacosa -- Ultraviolet irradiation of cells / David P. Turner, Anthony T. Yeung, and Alfonso Bellacosa -- Transient production of retroviral- and lentiviral-based vectors for the transduction of mammalian cells / Tiziana Tonini [and others].

Determination of functional viral titer by drug-resistance -- Colony assay, expression of green fluorescent protein, and '-galactoside staining / Tiziana Tonini [and others] -- Retroviral and lentiviral vector titration by the analysis of the activity of viral reverse transcriptase / Tiziana Tonini [and others] -- Single and double colloidal gold labeling in postembedding immunoelectron microscopy / Nicoletta Zini [and others] -- Multifluorescence labeling and colocalization analyses / Massimo Riccio [and others].

The dysregulation of the normal cell cycle underlies an enormous number of pathological conditions, including the neurodegenerative disorders, all cancers, and diabetes. In Cell Cycle Control and Dysregulation Protocols: Cyclins, Cyclin-Dependent Kinases, and Other Factors, expert laboratorians detail emerging methodologies for studying the cell cycle, its kinases, and kinase inhibitors. The authors focus on issues of gene expression in vivo and in vitro, the analysis of cyclin-dependent kinase inhibitors, protein degradation mediated by the proteosome, the analysis of the transformed cell phenotype, and innovative techniques to detect apoptosis. Many of the protocols are new, such as electron microsocopy to detect apoptosis and proteosome-mediated degradation, whereas others study the interactions between protein-protein, protein-DNA, and protein-RNA. The protocols follow the successful Methods in Molecular Biology™ series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. State-of-the-art and highly practical, Cell Cycle Control and Dysregulation Protocols: Cyclins, Cyclin-Dependent Kinases, and Other Factors offers pathbreaking researchers powerful tools for probing the cell cycle regulation across a wide variety of pathologies.

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