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Technique / Proteomics and protein biochemistry / Immunoprecipitation IP
| Immunoprecipitation (IP) is the technique of precipitating an antigen out of solution using an antibody specific to that antigen. This process can be used to enrich a given protein to some degree of purity. Co-immunoprecipitation (also known as a pull-down) can identify interacting proteins or protein complexes present in cell extracts: by precipitating one protein believed to be in a complex, additional members of the complex are captured as well and can be identified. The protein complexes, once bound to the specific antibody, are removed from the bulk solution by capture with an antibody-binding protein attached to a solid support. |
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Problem about the minimum amount of protein to do immunoprecipitation (Forum) Protein amount for immunoprecipitation Immunoprecipitation vs. Affinity Chromatography For Eventual Mass Spec Analysis (Forum) Methods to identify cellular proteins 
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Last update: 
11-May-2008 11:57 pm
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Chromatin Immunoprecipitation Assays: Application of ChIP-on-Chip for Defining Dynamic Transcriptional Mechanisms in Bone Cells. Methods Mol Biol. 2008;455:165-76
Authors: van der Deen M, Hassan MQ, Pratap J, Teplyuk NM, Young DW, Javed A, Zaidi SK, Lian JB, Montecino M, Stein JL, Stein GS, van Wijnen AJ
Normal cell growth and differentiation of bone cells requires the sequential expression of cell type specific genes to permit lineage specification and development of cellular phenotypes. Transcriptional activation and repression of distinct sets of genes support the anabolic functions of osteoblasts and the catabolic properties of osteoclasts. Furthermore, metastasis of tumors to the bone environment is controlled by transcriptional mechanisms. Insights into the transcriptional regulation of genes in bone cells may provide a conceptual basis for improved therapeutic approaches to treat bone fractures, genetic osteopathologies, and/or cancer metastases to bone. Chromatin immunoprecipitation (ChIP) is a powerful technique to establish in vivo binding of transcription factors to the promoters of genes that are either activated or repressed in bone cells. Combining ChIP with genomic microarray analysis, colloquially referred to as "ChIP-on-chip," has become a valuable method for analysis of endogenous protein/DNA interactions. This technique permits assessment of chromosomal binding sites for transcription factors or the location of histone modifications at a genomic scale. This chapter discusses protocols for performing chromatin immunoprecipitation experiments, with a focus on ChIP-on-chip analysis. The information presented is based on the authors' experience with defining interactions of Runt-related (RUNX) transcription factors with bone-related genes within the context of the native nucleosomal organization of intact osteoblastic cells.
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