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Protocol
In Situ Nuclear Organization of Regulatory Machinery
Regulatory machinery for gene expression, replication, and repair are architecturally organized in nuclear microenvironments. This compartmentalization provides threshold concentrations of macromolecules for t...
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Protocol
In Situ Immunofluorescence Analysis
Immunofluorescence is one of the most widely used techniques to study the localization of transcription factors, proteins, and structural components of nuclear architecture and cytoarchitecture. High-resolution i...
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Protocol
In Situ Immunofluorescence Analysis
This technique is used to visualize sites of active transcription in a permeabilized cell and does not require radiolabeled molecules (e.g., [3H] Uridine). Nonradioactive ribonucleic acid (RNA) precursors (e.g., ...
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Protocol
Immunofluorescence Analysis Using Epitope-Tagged Proteins
The resolution that can be attained in assessment of the intranuclear localization of cellular proteins is dependent on specificity of the antibodies. Primary antibodies should be well characterized. We recomm...
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Protocol
Analysis of In Vivo Gene Expression Using Epitope-Tagged Proteins
Only a handful of tagged molecules have been used successfully in vivo to monitor gene expression. The most commonly used are β-galactosidase (β-gal) and green fluorescent protein (GFP). Both molecules pose li...
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Protocol
Chromatin Immunoprecipitation
Fundamental parameters of biological control require sequence-specific protein-deoxyribonucleic acid (DNA) interactions. The packaging of DNA into higher order chromatin controls accessibility to DNA binding p...
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Protocol
Protein-Deoxyribonucleic Acid Interactions Linked to Gene Expression
The electrophoretic mobility shift assay (EMSA) determines sequence-specific protein-deoxyribonucleic acid (DNA) interactions (e.g., transcription factors with cognate regulatory elements). This assay is based...
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Protocol
Protein-Deoxyribonucleic Acid Interactions Linked to Gene Expression
Map** DNase I hypersensitive sites can often localize the control regions in a eukaryotic gene. It is generally believed that chromosomal regions with loose or more open conformation are sensitive to DNase I...
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Protocol
Protein-Deoxyribonucleic Acid Interactions Linked to Gene Expression
Ligation-mediated polymerase chain reaction (LM-PCR) is used for high-resolution genomic footprinting. This technique was originally developed to study in vivo protein-deoxyribonucleic acid (DNA) interactions ...
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Protocol
Stimulation of Anti-HIV-1 Cytotoxic T Lymphocytes by Dendritic Cells
Dendritic cells (DC) are highly specialized antigen-presenting cells (APC) in both primary and secondary T-cell responses. This may be related to their expression of high levels of MHC class I and II antigens (1-
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Protocol
Methods for Chemoprotection and Chemosensitization
Surgery, radiation therapy, and chemotherapy have been applied to the curative therapy of 50% of cancer patients in the United States during the past 100 years. It is clear that the chemotherapeutic agents use...
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Protocol
Dynamic and Exchange Processes in Macromolecules Studied by NMR Spectroscopy
It is normal for a biological system to be in a dynamic state. The function of many biological systems depends on their flexibility, and here NMR can provide the experimental basis for investigating the mechan...
