[RASMB] Fluorophore with Interference Optics

[Matthew Hockin]

Hi Rob-

Its been a while since I have been on here and even longer since I have
worked with these instruments regularly, so perhaps we can take my answer
as a bump to your post -

That said, interference optics are "seeing" a change in refractive index
between the two sides of the cell, although I presume FITC would be
different than an amino acid with respect to its concentration effect
contribution to the observed signal its probably not a big change if its
part of a much bigger protein, and in any case is attached to the protein,
so any (possibly- slight- I assume) effect it might have on the
interference signal would only show up if your trying to quantitate the
protein concentration.  Since its attached to the protein I would begin
with the assumption that it will not affect the outcome in terms of the
calculated S values etc, beyond whatever effect it has on protein behavior
as a macromolecule.

I will await further discussion as a refresher.  I know I have done this in
the past but cannot recall if we observed fringe offsets, I don't think we
did, as we were looking at larger proteins and a single FITC would not be a
major change, if your working with really small stuff maybe its different,
or I may be recalling incorrectly.

On Tue, Jun 2, 2015 at 2:22 PM, Robert Shaffer <rshaff at bu.edu> wrote:

> If I use a FITC-labeled ligand with my protein, will the fluorophore cause
> any issue with the interference optics for a sedimentation velocity run I
> plan to do next week? Or, does fluorescein not affect the quality of
> interference data?
>
> Thank you,
> Rob
>
> --
> Robert Shaffer
> rshaff at bu.edu
> MCBB, PhD Program
> Boston University
>
>
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