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Reagent Preparation:
Dissolve Indo-1 (Molecular Probes catalog No. I1226) in DMSO to a final concentration of 2 mg/ml (25 ul of DMSO into 50 ug of Indo-1).
Mix quantities of probenecid (Sigma Catalog No. P8761) and water to yield a 100 mM solution. Weigh 285 mg of probenecid, add 8 ml of dH2O, then add 1 M NaOH until dissolved. The pH should be between 9 and 10. Store for 1 month at room temperature.
Dissolve Ionomycin (Calbiochem Catalog No. 407950) in 100% ethanol for a final concentration of 1 mg/ml. Store at –20°C. This is used as a control for calcium flux.
Cell Loading Media (CLM) is Hanks Balanced Salt Solution (HBSS) containing 1 mM calcium, 1 mM magnesium, 1% FBS or 0.5% BSA and can be stored up to 1 month at 4°C.