@article{87516,
  keywords = {Animals, Drosophila, Chromatin Immunoprecipitation, DNA-Binding Proteins, Embryo, Nonmammalian, Cross-Linking Reagents, Tissue Fixation, Drosophila Proteins, Succinimides},
  author = {Tsutomu Aoki and Daniel Wolle and Ella Preger-Ben Noon and Qi Dai and Eric Lai and Paul Schedl},
  title = {Bi-functional cross-linking reagents efficiently capture protein-DNA complexes in Drosophila embryos.},
  abstract = { Chromatin immunoprecipitation (ChIP) is widely used for mapping DNA-protein interactions across eukaryotic genomes in cells, tissues or even whole organisms. Critical to this procedure is the efficient cross-linking of chromatin-associated proteins to DNA sequences that are in close proximity. Since the mid-nineties formaldehyde fixation has been the method of choice. However, some protein-DNA complexes cannot be successfully captured for ChIP using formaldehyde. One such formaldehyde refractory complex is the developmentally regulated insulator factor, Elba. Here we describe a new embryo fixation procedure using the bi-functional cross-linking reagents DSG (disuccinimidyl glutarate) and DSP (dithiobis[succinimidyl propionate). We show that unlike standard formaldehyde fixation protocols, it is possible to capture Elba association with insulator elements in 2-5 h embryos using this new cross-linking procedure. We show that this new cross-linking procedure can also be applied to localize nuclear proteins that are amenable to ChIP using standard formaldehyde cross-linking protocols, and that in the cases tested the enrichment was generally superior to that achieved using formaldehyde cross-linking. },
  year = {2014},
  journal = {Fly (Austin)},
  volume = {8},
  pages = {43-51},
  issn = {1933-6942},
  doi = {10.4161/fly.26805},
  language = {eng},
}