Drying down Cy3-NHS into aliquots

[lechristophe]

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Hi,

A question for the people who make their fluorescent probes:

I have split 1 mg Cy3-NHS into 10 x 0.1 mg using 20 µL DMSO into 10 x 2 µL.
Now I'm trying to evaporate the DMSO but it seems quite difficult (it's
been two hours in the Speedvac/cold trap and it's still not dried). Any
suggestions on how to proceed (or an alternative method to DMSO)?

Thanks for your help,

Christophe

[Peter O'Toole]

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Hi Christoph

I used to bring Cy3 up in degassed dH2O (with argon), aliquot in to
eppendorf tubes and capped under a flush of argon and plunge the tube in
to liquid nitrogen. I would then store in liquid nitrogen until
needed.   It worked well and gave reproducibe labelling ratio's.

Best

Pete

On 18/09/2014 12:29, Christophe Leterrier wrote:

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> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi,
>
> A question for the people who make their fluorescent probes:
>
> I have split 1 mg Cy3-NHS into 10 x 0.1 mg using 20 µL DMSO into 10 x 2 µL.
> Now I'm trying to evaporate the DMSO but it seems quite difficult (it's
> been two hours in the Speedvac/cold trap and it's still not dried). Any
> suggestions on how to proceed (or an alternative method to DMSO)?
>
> Thanks for your help,
>
> Christophe

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[Iain Johnson]

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DMSO is very hard to evaporate (boiling point 189C). Use ethyl acetate instead as the solvent vector. Much more volatile and won't cause ester hydrolysis, unlike water, methanol etc.

Iain

Sent from my iPhone

On Sep 18, 2014, at 4:29 AM, Christophe Leterrier <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi,
>
> A question for the people who make their fluorescent probes:
>
> I have split 1 mg Cy3-NHS into 10 x 0.1 mg using 20 µL DMSO into 10 x 2 µL.
> Now I'm trying to evaporate the DMSO but it seems quite difficult (it's
> been two hours in the Speedvac/cold trap and it's still not dried). Any
> suggestions on how to proceed (or an alternative method to DMSO)?
>
> Thanks for your help,
>
> Christophe