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> I was wondering if anyone has experience loading COS-7 cells with
> Indo for calcium mobilization analysis. We've been having a tough
> time with this. The Indo seems to be ok, since we've loaded other cells
...etc.
> Peter Lopez
I asked a similar question recently regarding CHO cells
and one of the suggestions from the list was to use
verapamil. We tried loading in 4uM INDO-1, 30uM verapamil
for 1 hour (including the pluronic) and it worked great.
The fluorescence we got was an order of magnitude better
than loading in the absence of verapamil. I would suggest
this as a first try to see if the COS cells are actively
pumping out the dye as it is being cleaved. Good luck!
Keith
**********************************************************************
*
Keith A. Kelley kelley@wh.bayer.com *
Institute for Research Technologies * "No tree is too big
Bayer Corporation * for a short dog to
400 Morgan Lane Ph 203-931-5303 * lift his leg on"
West Haven CT 06516 FAX 203-937-5467 *
* Texas Bix Bender
**********************************************************************
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web