during a conversation with some chemists, I´ve been told that formaldehyde reversing X-linking (high temperature and high salt concentration) may result in a loss of histone acetylation levels due to some similarities between the nature of bond that formaldehyde and the acetyl-group have. In other words, reverse X-linking might determine a sort of "chemical deacetylation".
Is anybody aware of such issue?
I´ve been working on ChIP for a long time and this issue doesn´t really matter as the IP is already done at the stage of reverse X-linking, but now, since I´m planning to perform a specific assay which is not a conventional ChIP, such issue might be critical to be addressed.
Basically, what I´m planning to do is a sort of Co-IP assay, by pulling down of a specific histone modification and then performing a western blot detection against acetylated residues. Since I think I need either to perform X-linking to keep the complexes or to perform (by mere assumption) reverse X-linking to properly run a PAGE, I´m wondering wheather the reverse X-linking itself might somehow affect the outcome by "chemically deacetylating" my targets.
I´m sorry for the quite weird question and I hope to read your feedback.
Edited by HP1b, 23 June 2009 - 02:18 AM.