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That sounds like the Roger Tsein tetracystein tag approach?  Originally
published in Science a while back.

I've heard a couple of recent talks that use this, with bicistronic
fluorescent probes, and fluorescein. 488nm light then inactivates the
protein, as the excited fluorescein results in release of free radicals.

So, depends what you want to use it for.  Bleaching may not be a good idea -
unless of course you want to inactivate your protein?

Michelle


On 21/2/05 3:05 pm, "Alistair Barber" <[log in to unmask]> wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Does anyone have experience using the Lumio construct from Invitrogen
> in timelapse and/or FRAP confocal studies?
> Lumio is a six amino acid sequence that can be used to tag proteins in
> the same way as GFP, but is less likely to interfere with normal
> peptide function because of its small size. The construct only becomes
> fluorescent when the transfected cells are exposed to a cell-permeable
> substrate. What is not clear to me from the Invitrogen literature is if
> Lumio can be bleached in the same way as GFP, or if there will be rapid
> recovery while the substrate is still present in the cell?
>
> -Alistair Barber

Dr Michelle Peckham
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School of Biomedical Sciences
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