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> Along with several other cell lines, CHO cells can express very high levels
> of p-glycoprotein, and this actively pumps indo-1 out of the cells. The
> solution is to load in the presence of a pgp inhibitor such as 10-30 uM
> verapamil. We published an account of this problem in Cytometry last year.
That is all OK, but if one wants to use INDO-1 as calcium signal detector
(as it is mostly used for), the simultaneous use of verapamil (or another
calcium channel blocker) to block P-glycoprotein may not be the best
idea. I'd suggest reserpine (titrate first, starting from 1 microgram per
ml). It worked ok in our hands and did not affect calcium signal,
although it might be nasty if you want to culture the cells for a longer
time afterwards.
Jacek
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Jacek M. Witkowski
Department of Histology
Medical University of Gdansk, Poland.
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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;
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