Mark, We saw unexpected results looking at calcium flux in Jurkat. Forcing the culture to expand a neomycin resistant clone had resulted in a drug resistant (MDR phenotype) cell line. If you have the same, your staurosporin treatment could indirectly block the efflux of R123, thus causing a net increase in dye retention. MAK. Mark Cozens wrote: > I'm studying honours in the Dept. Pathology and am looking at > depolarisation of the mitochondrial membrane as an early event in > apoptosis of Jurkat T cells. I'm staining cells with the > mitochondrial specific dye Rh123 and am using 5microM staurosporine > treated cells as a positve control to show mitochondrial > depolarisation. However, staurosporine treatment appears to be > causing an increased uptake of Rh123. I'm wondering if this is due to > mitochondrial changes or another property of staurosporine treatment > that is increasing uptake of the dye. > If anyone has any experience with this area or any ideas, feedback > would be muchly appreciated. > Thanks, > Anna Hearps > (ahearps@postoffice.utas.edu.au) > -- > Mark A Cozens > Division of Pathology > University of Tasmania > ph 03 62264828 > fax 03 62264833 -- Mark A. KuKuruga, Managing Director University of Michigan Core Flow Cytometry <http://www.cancer.med.umich.edu/flow_cytometry> phone: 734-647-3216 fax: 734-936-7376 kukuru@umich.edu
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