Debate

Tom Anderson via methods%40net.bio.net (by ucgatan from ucl.ac.uk)
Tue Mar 4 12:01:47 EST 2008


On Fri, 29 Feb 2008, DK wrote:

> In article <Pine.GSO.4.58.0802291108450.8100 from socrates-a.ucl.ac.uk>, Tom Anderson <ucgatan from ucl.ac.uk> wrote:
>
> >Note that in the case of some fluorescent proteins, there's also a risk of
> >oligomerisation artifacts: wildtype FPs (all or just some) are tetrameric
>
> Happily, not the classic Aequorea GFP. That one only forms weak dimers
> that are detectable only at high concentrations.  Even in many crystals
> such a dimer is not observed.

Ah! I didn't realise that, thanks and apologies.

> >There is a relatively new alternative to GFP for fluorescent tagging of
> >proteins in live cells in the shape of the FlAsH/ReAsH system:
> >
> >http://www.invitrogen.com/content.cfm?pageid=11922
> >
> >Basically, you put a magic tag on your protein,
>
> The magic tag with two exposed Cys is a subject to many crosslinking
> artifacts. Probably not a problem for most cytoplasmic proteins
> (reducing environment) but must be a huge problem for anything that goes
> through secretion pathway.

Yes, i assume it'd be a complete non-starter for anyhing outside the
cytoplasm.

tom

-- 
Tom Anderson, MRC Laboratory for Molecular Cell Biology, UCL, London WC1E 6BT
(t) +44 (20) 76797264   (f) +44 (20) 76797805   (e) thomas.anderson from ucl.ac.uk


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