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We make a stock 1 mM solution in absolute ethanol and keep it at -20oC
for months. Working dilution is prepared immediately before staining in
appropriate buffer - usually PBS supplemented with 5mM glucose or HBSS.
Working concentration depends on the actual cells and is usually within
the 1-25uM range. If you want simultaneous surface phenotyping, use as
low R123 conc as possible - you will need heavy compensation esp. using
PE. RED613- or RED670- coniugates may be better for this purpose.
You may want to look at our paper in J. Immunol. 1993, 150:1296 for some
methods details. Good luck!
Jacek
On Thu, 25 Jan 1996, Glenda, M, Davison wrote:
> Hello from Cape Town,
>
> Can anyone tell me what diluent to use when making up rhodamine 123
> and also how to store it once it has been diluted. We would like to
> use rhodamine 123 to detect the presence of the p-glycoprotein in
> leukaemic cells.
>
> Many thanks in advance,
>
> Glenda.
>
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