Titlebook: Cell Cycle Control and Dysregulation Protocols; Antonio Giordano,Gaetano Romano Book 2004 Humana Press 2004

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Immunofluorescence Analysisine). Nonradioactive ribonucleic acid (RNA) precursors (e.g., 5-bromouridine-5′-triphosphate [BrUTP]) are used and can be detected by using fluorescently labeled antibodies. Procedures for BrUTP of labeling transcription sites require manipulations that are best applied to adherent cells but can be

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Analysis of In Vivo Gene Expression Using Epitope-Tagged Proteinsand green fluorescent protein (GFP). Both molecules pose limitations with in vivo detection. When using GFP, autofluorescence may be encountered in tissues and tissue sections. Similarly, when using β-gal as a marker, the penetration of X-gal substrate is difficult in tissues with extensive extra ce

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Protein-Deoxyribonucleic Acid Interactions Linked to Gene Expressiontors with cognate regulatory elements). This assay is based on reduced electrophoretic mobilities of protein/DNA complexes through a nondenaturing polyacrylamide gel compared with unbound DNA fragments or double-stranded oligonucleotides. If multiprotein complexes bind, the migration rate slows furt

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Protein-Deoxyribonucleic Acid Interactions Linked to Gene Expressionth loose or more open conformation are sensitive to DNase I cleavage. By comparing the patterns of hypersensitive sites obtained by digestion of silent genes with those obtained when the genes are actively transcribed, regions that participate in gene regulation (either induction or suppression) can

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Protein-Deoxyribonucleic Acid Interactions Linked to Gene Expressiony in vivo protein-deoxyribonucleic acid (DNA) interactions at regions of genes important for transcriptional regulation. Successful analysis of in vivo occupancy of gene regulatory elements by LM-PCR largely depends on the strategy and quality of reagents. The experimental outline for LM-PCR reactio

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Assays for Cyclin-Dependent Kinase Inhibitorse if associated with its cyclin partner and if the complex is phosphorylated at specific activating residues (threonine 160/161; .. and .). The progression through the cell cycle is mediated by the sequential activation of CDKs during different phases of the cycle. The G1/S phase transition of the c