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Date: | Mon, 1 Dec 2014 11:31:58 -0600 |
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I haven’t tried them yet, but most any cell permeant reagent has to contend with the powerful xenobiotic export mechanisms of Dicty. Basically, even if the tags are properly expressed in the cells, finding the appropriate cell permeant version to be present in high enough concentrations internally can be an issue once the cells are brought up to physiological temperatures. If the labeling bonds can happen at the cold, may be better, but I think that is a different set of kinetics.
> On Dec 1, 2014, at 11:03 AM, Robert Insall <[log in to unmask]> wrote:
>
> Dear Dictyophiles,
>
> Has anyone ever tried using either HALO or SNAP tags in Dicty? They offer the promise of brighter, more photostable dyes and smaller fusion partners than GFP, but I wonder if the cell-permeant ligands will get into Dicty cells well enough, or if there’s a chemical background that interferes.
>
> Both tags also allow affinity purification from lysates, which is an added incentive.
>
> Robert
>
> --------------------------------------
> Professor Robert Insall
> CR-UK Beatson Institute for Cancer Research
> Switchback Road, Bearsden
> Glasgow G61 1BD, UK
>
> Tel. (0/44) 141 330 4005
> Web http://www.beatson.gla.ac.uk/robert_insall
> --------------------------------------
>
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