Re: Ca-indo and gfp?

From: Eric Van Buren (aa9080@wayne.edu)
Date: Fri Jan 25 2002 - 17:52:10 EST


Andreas,

I believe all the "green" GFP variants are UV-excitable. Is the GFP
fluorescence showing up in the Indo-1 "blue (long)" detector (~525BP)?
Perhaps you could try DsRed instead of GFP.

Does your system properly measure calcium responses with other cells?

I have successfully run indo-1 samples on a FACS Vantage, and indo-1
with PE, but I have not tried indo-1 with GFP.

Good luck,
Eric


>Hi everybody,
>we have a BD FACS-Vantage and have tried to measure
>Ca-fluxes in GFP overexpressing cells. The cells used are
>a low-adherent tumor cell line and were detached by
>little mechanical force (tapping the flask). We used INDO
>as the Ca-dependent fluorescent dye.
>Unfortenately, we had bad luck. Even with ionomycin, we did
>not get a Ca response.
>Has somebody experience with this combination (GFP INDO)
>and can help us?
>
>A second question:
>Who knows a flow cytometry company working about blood
>homeostasis (platelet activation ...) in Europe?
>
>Thanks in advance
>
>Andreas
>
>
>
>PD Dr. Andreas Simm
>Universitaet Halle Wittenberg
>Klinik fuer Herz- und Thoraxchirurgie
>Ernst-Grube Str. 40
>D-06120 Halle
>
>Tel.: +49 (0) 345 557 2647
>                  552 2878
>FAX:  +49 (0) 345 557 2782
>                  552 2890


Eric Van Buren, aa9080@wayne.edu
Karmanos Cancer Institute and Immunology & Microbiology
Wayne State University, Detroit, Michigan, USA



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