Hi Greg,
A number of your questions (and others) have been addressed in an excellent
article in a new journal:
Tonge R. et al. (2001) Proteomics 1 pp. 377-396 (I think you can download it
for free from their web site)
However, as Jeff mentioned, there are a number of caveats in the use of this
technology.
Good luck
Axel
"Jeffrey A Kowalak, Ph.D." wrote:
> Greg,
>
> I have no direct experience with Cy3/Cy5 but my understanding is that only
> 1-2% of the proteins get labeled. Apparently, this is enough for fluorescent
> visualization. Excess reagent would be removed during electrophoresis
> (assuming there is also some kind of quenching step). At 1-2% labeling,
> there should be no problem with downstream MS. WARNING: Imaging system is a
> major expense ~$60,000 USD.
>
> Regards,
> Jeff
>
> Jeffrey A Kowalak, Ph.D.
> Mass Spectrometry Facility Director
> Laboratory of Neurotoxicology
> National Institute of Mental Health
> Building 10 Room 3D42
> Bethesda, MD 20892-1262
>
> Telephone: 301-496-4242
> Facsimile: 301-480-0198
> e-mail: jkowalak@intra.nimh.nih.gov
>
> -----Original Message-----
> From: Association of Biomolecular Resource Facilities
> [mailto:abrf-request@aecom.yu.edu]On Behalf Of Greg Grant
> Sent: Monday, April 16, 2001 1:10 PM
> To: Recipients of ABRF List
> Subject:
>
> Dear Colleagues,
>
> Can anyone provide any information on the Cy 3 and Cy 5 dyes being
> marketed by Amersham as to their properties and compatability with mass
> spec analysis. Like, how do they interact with proteins? How sensitive are
> they? Do they need to be removed prior to analysis? and anything else.
>
> Thanks,
>
> Gregory A. Grant
> ggrant@pcg.wustl.edu
> 314-362-3367
> FAX 314-362-4698
--------------------------------------------------------------------------
Axel Ducret, Ph.D. Senior Research Biologist Merck-Frosst Canada Inc. Dept. Biochemistry and Molecular Biology P.O. Box 1005 Pointe-Claire-Dorval PQ H9R 4P8 Canada
tel. + (514) 428-3428 fax + (514) 428-4900
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