Abstract
This is a protocol of Ca2+ imaging experiment using Ca2+ indicator Fura-2. Ca2+ imaging is an efficient and quantitative method for measuring cytosolic and internal store Ca2+ levels, as well as their dynamic changes.
Materials and Reagents
Equipment
Procedure
Note: Target cells: concentration of Fura-2-AM may need to be optimized depending on cell types to be measured. The following protocols are designed for HEK293 cells.
Recipes
References
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You can use 2 μM thapsigargin (TG) or 2 μM Ionomycin, which depletes ER Ca2+ store and induces Ca2+ influx.
Hope the answer posted below have answered your question.
GFP should have minor effects on Fura-2 measurement, because its major excitation peak is over 450nm, while Fura-2 is excited at 340 and 380nm. But a GFP-only control would be necessary for the measurement.Fura-2(Calcium free) and Fura-2(Calcium bound) have different excitation wavelength. When Fura-2 is bound to calcium, its excitation peak is 340nm. When Fura-2 is not bound to calcium, it will be 380nm.