Re: FITC unconjugated

From: Mario Roederer (Roederer@drmr.com)
Date: Fri Aug 06 1999 - 13:17:28 EST


>Hello,
>Does anyone have any experience with the storage of unconjugated FITC
>(isomer 1, Sigma, F7250) ? I have dissolved it in PBS, pH8, 0.1mg/ml
>and I keep it in the dark at 3-4°C.  Does anyone know something about
>the stability in this form and how long it can be stored like this ?
>
>Thanks
>Isabel

About 30 minutes.  FITC reacts with water quite rapidly...  never store it
in water!

Since the cost of FITC is so low compared to the cost of everything else
associated with the reaction, you should always use a freshly-opened vial
when you are about to use it; discard the rest.  Also, dissolve the FITC in
DMSO and then add it to your reaction.  Make sure that your DMSO is the
highest-grade possible, and keep the DMSO in a dessicator to make sure it
has no water.

Note that you cannot use pipetteman to accurately pipet DMSO--you have to
use glass (not plastic!) pipettes.  Typically, we break the ampule, and add
1.0 mL of DMSO from a glass pipet.  This results in 10mg/ml solution, which
is great for most reactions.  (Then you will have to add the small amount
you need using a pipettman; using yellow tips works reasonably well but you
have to operate quickly:  the DMSO does not hold well inside the tip and
leaks out).

FITC in DMSO is relatively stable (i.e., hours... not days), when kept
unexposed to air & water.

mr



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