Indo-1 loading

kelley@wh.bayer.com
Thu, 21 Dec 95 10:25:25 -0500

> Hello

> 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
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Keith A. Kelley kelley@wh.bayer.com *
Institute for Research Technologies * "No tree is too big
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400 Morgan Lane Ph 203-931-5303 * lift his leg on"
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