calcium flux in leishmania

Barbara Butcher (Barbara_Butcher@d4.niaid.pc.niaid.nih.gov)
Wed, 16 Aug 95 09:52:26 EST

i've been trying to look at calcium flux in leishmania major promastigotes
with various indicators (fluo-3, calcium green, fura red, indo-1) and have
had no success generating a positive response using the ionophores A23187,
oligomycin and ionomycin. i've been able to load calcein-am into the
parasites and detect the fluorescence, so i know that the cells have
esterase activity. i've tried verapamil to inhibit efflux of the dye, i've
loaded the parasites at 20C and 4C (they grow at 26C), i've tried loading
with and without pluronic f-127, with and without serum, with and without
1.5% sucrose. nothing has worked thus far!! can anyone help before i take
a job at mcdonald's?

thanks in advance,

barbara butcher
lab of parasitic diseases
niaid/nih
bethesda, md 20892
301 496-0577 (phone)
301 402-0890 (fax)