SpringerProtocols: Abstract: Chromatin Immunoprecipitation for Studying Transcriptional Regulation in Xenopus Oocytes and Tadpoles

Chromatin Immunoprecipitation for Studying Transcriptional Regulation in Xenopus Oocytes and Tadpoles

By: David Stewart², Akihiro Tomita³, Yun-Bo Shi³, Jiemin Wong²

Abstract

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Understanding the accurate temporal and spatial regulation of gene expression during development requires knowledge of the spectrum of transcription factors and cofactors involved and their functional interplay with chromatin. Chromatin immunoprecipitation (ChIP) has become a powerful technique that allows us to do so. A typical ChIP assay involves (1) treating cells or tissues with formaldehyde to rapidly crosslink chromatin-associated proteins to DNA, (2) shearing chromatin by sonication into small fragments, (3) immunoprecipitation of the proteins of interest, (4) reversal of crosslinking, and (5) quantitating the specific associated DNA sequences by PCR. Here we present and discuss the protocols we have developed over the years for ChIP assays using Xenopus oocytes and tadpole tissues as experimental materials.

Affiliation(s): (2) Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX
(3) Section on Molecular Morphogenesis, Laboratory of Gene Regulation and Development, National Institute of Child Health and Human Disease, National Institutes of Health, Bethesda, MD

Book Title: Xenopus Protocols: Cell Biology and Signal Transduction

Series: Methods in Molecular Biology | Volume: 322 | Pub. Date: Dec-13-2005 | Page Range: 165-181 | DOI: 10.1007/978-1-59745-000-3_12

Subject: Cell Biology

Key Words: Chromatin immunoprecipitation (ChIP) - tadpole - thyroid hormone receptor - transcription - Xenopus oocyte

References

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