Special Issue Techniques
Chromatin immunoprecipitation in early Xenopus laevis embryos
Article first published online: 30 MAR 2009
Copyright © 2009 Wiley-Liss, Inc.
Special Issue: Special Focus on Xenopus
Volume 238, Issue 6, pages 1422–1432, June 2009
How to Cite
Blythe, S. A., Reid, C. D., Kessler, D. S. and Klein, P. S. (2009), Chromatin immunoprecipitation in early Xenopus laevis embryos. Dev. Dyn., 238: 1422–1432. doi: 10.1002/dvdy.21931
- Issue published online: 12 MAY 2009
- Article first published online: 30 MAR 2009
- Manuscript Accepted: 22 FEB 2009
- National Institutes of Health. Grant Numbers: T32-GM007229, T32-HD007516, R01-GM64768, R01-GM76621
- National Sciences Foundation. Grant Number: IOS-718961
- chromatin immunoprecipitation (ChIP);
- transcription factor;
Chromatin immunoprecipitation (ChIP) is a powerful method for analyzing the interaction of regulatory proteins with genomic loci, but has been difficult to apply to studies on early embryos due to the limiting amount of genomic material in these samples. Here, we present a comprehensive technique for performing ChIP on blastula and gastrula stage Xenopus embryos. We also describe methods for optimizing crosslinking and chromatin shearing, verifying antibody specificity, maximizing PCR sensitivity, and quantifying PCR results, allowing for the use of as few as 50 early blastula stage embryos (approximately 5×104 cells) per experimental condition. Finally, we demonstrate the predicted binding of endogenous β-catenin to the nodal-related 6 promoter, binding of tagged Fast-1/FoxH1 to the goosecoid promoter, and binding of tagged Tcf3 to the siamois and nodal-related 6 promoters as examples of the potential application of ChIP to embryological investigations. Developmental Dynamics 238:1422–1432, 2009. © 2009 Wiley-Liss, Inc.