Sedimentation and Immunoprecipitation Assays for Analyzing Complexes that Repress Transcription
Co-repressor proteins function as platforms for the assembly of multi-subunit complexes that mediate transcriptional repression. Common components of such complexes are histone deacetylases, which catalyze the removal of acetyl groups from the tails of histones within nucleosomes, resulting in chromatin compaction and gene repression. In addition, co-repressor complexes generally interact with sequence-specific DNA-binding proteins that direct association with regulatory elements in the genome. Thus, identifying proteins that stably associate with co-repressors can provide insights regarding the biochemical function and target gene specificity of these molecules. Here, we describe a density gradient fractionation method for determining whether a co-repressor is incorporated into high-molecular complexes within cells and for identifying potential constituents of these complexes. We also describe a co-immunoprecipitation assay for confirming and further studying interactions between co-repressors and other proteins that may represent functional partners.
- PC/GENE: Searches for Functional Sites in Nucleic Acids and Proteins
- Quantitative Stem-Loop RT-PCR for Detection of MicroRNAs
- PCR In situ hybridization
- Quantification of Methylated DNA by HeavyMethyl Duplex PCR
- Experimental Designs for QTL Fine Mapping in Rodents
- Forensic DNA-Typing Technologies: A Review
- Computational Identification of sRNA Targets
- Novel Methodology for Immobilization of Biomolecules on the Surface of a Photoresponsible Polymer Containing Azobenzene Moiety
- The Micronucleus Assay Determination of Chromosomal Level DNA Damage
- Genomic Analysis of Transgenic Animals by the Polymerase Chain Reaction