(by peterh from my.molbio.ku.dk)
Thu Nov 22 12:17:44 EST 2007
Thank you for your great answers.
I realize that achieving the actual chemistry of crosslinking two carboxyl groups is a lot tougher than I anticipated.
If I got it right, using a carbodiimide (like EDC) will be a problem for me because it prepares carboxyls to react with amines and thereby Proteins with DNA. Adding high concentrations of polyamines might out-compete DNA-amines but might also cause very little crosslinking since carboxyl groups have to bind the same polyamine-molecule to be crosslinked.
The use of more target-specifik crosslinking methods (like transglutaminase under a controlled promoter) seems less attractive to me because I want the crosslinking to be fairly unspecifik and capable of crossbinding lots of different proteins.
However, knowing that I have to rethink my strategy is also of great value. :o)
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