calcium flux

From: gattal@infos1.casaccia.enea.it
Date: Wed Apr 21 1999 - 10:35:23 EST 




Greetings,

Is anyone aware of methods to measure intracellular calcium flux with
fluorometer? 

I'm interested in monitoring influx and efflux in cytoplasm of freshly
isolated murin T CD4+ cells.

My protocol includes use of Fluo-3 in HBSS with 650microM CaCl2 and
Glucose for 45 min, 3 washes, and measure at lambda ex. 480 nm and
lambda em. 520 nm. The cells show Ca2+ influx across plasma membrane in
presence of CaCl2 in the medium after EGTA addition, but show no
intracellular Ca2+ pool mobilisation ( I have no response to
thapsigargin). Do you know if the buffer I'm using could empty cells
stores? For example Mg could be important or CaCl2 is too low? Do you
have any other good ideas for my problem? 

Thanks in advance



Lucia Gatta



Laboratorio di Immunologia


Università di Roma Tor Vergata


V. Della Ricerca Scientifica


00133 Roma


TEL +39-6-72594242

FAX +39-6-2023500













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