Does anybody know the half-life of formaldehyde protein-protein, protein-RNA and protein-DNA crosslinks when the sample is incubated at 37C?
I read somewhere that tris and other primary amine buffers are able to partially reverse formaldehyde crosslinking. However, aldehydes can crosslinks proteins through more than one reaction and some are irreversible.
I don't think they would reverse crosslinking, but they are inhibitors because they compete with the amino groups in proteins
Hi Tomas. Formaldehyde X-linking can be reversed, but apparently there is some disagreement in the litterature on whether primary amines are required for this.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3077733/pdf/main.pdf
Heat will reverse the cross-links. This is most efficiently done at 70C for 30 or 45 minutes (depending which publication you agree with). I was wondering how fast this process is at 37C.
Steingrimur, in the article you linked, formaldehyde cross-links are reversed by an overnight incubation at 42C. This is the closest to 37C I've found as of yet.
Figures of crosslinked samples incubated at 37C for 10 minutes show no reversal;
http://jb.asm.org/content/187/18/6536.figures-only
Hi Sebastian. Are you trying to reverse or protect formaldehyde X-linking at 37oC?
I don't know if these will help:
Methods for isolating covalent DNA–protein crosslinks.
http://toxicology.usu.edu/endnote/DNA-protein-crosslinksIsolationMethod-AnalBiochem2005.pdf
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2771946/pdf/nihms90840.pdf
Thanks for contributing these papers and for the help. It seems that in the second one, cross links are reversed by an overnight incubation at 68C. I know from two other papers that crosslinks are most efficiently reversed by an incubation of 45 minutes at 70C. Unfortunately, this doesn't indicate the stability of these crosslinks at 37C. The paper I link to in this comment does however show that formaldehyde crosslinks between DNA and histones have a half-life of 18.1 at 37C. This is probably the closest I can get to a real half-life for my own samples.
http://carcin.oxfordjournals.org/content/21/8/1573
Hey there, table 1 in this article from 2014 measures the half-life of the crosslink at 37 degrees as 22.7 h.
Cheers,
Ruben
https://www.ncbi.nlm.nih.gov/pubmed/24848408
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