Which cross-linking reagent?
happytobe at home.eu
Sun Feb 10 10:31:33 EST 2002
"Ricky Boernke" <ricky_boernke at gmx.net> a écrit dans le message news:
3C57DF20.301741E2 at gmx.net...
> Dear Newsgroup,
> we would like to cross-link a bait protein expressed in
> Arabidopsis to potential binding partners and subsequently
> purify the complex using our protein as a tag.
> However, we are quiete unsure which cross-linking reagent
> we should use for this purpose. The Pierce catalogue offers
> quiete a few different types and I would suppose they are all
> suited for different purposes. Is there anything like a
> guide to protein cross-linking? I couldn't find anything more
> general. If anybody could point me into the right direction?
I remember some nice litterature on histone chemical crosslinking, as well
as an old but nice review on glutaraldehyde crosslinking (Ann Rev
Biochemistry, 1977 ?)
As far as I tried (and this was painful) , the rule is : use more than one
crosslinker, since no complex stabilization may just mean that the spacer
between the crosslinker reactant groups is just too short; a too long
spacer, of course, will stabilize non specific complexes :)
Agree with Kyle's advice !
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