Cell-permeable, green fluorescent calcium indicator dye (Kd = 0.35µM), ideal for calcium imaging (e.g. live-cell imaging and intracellular calcium flux detection). Non-ratiometric. Widely used in fluorescence microscopy, flow cytometry, and high-throughput calcium assays. Brighter, quicker to penetrate cells, more stable and shows higher affinity for Ca2+ than Fluo-3. Upon binding calcium, exhibits increased fluorescence in response to 488nm excitation, making it an excellent choice for studying calcium signaling in neurons, cardiomyocytes, other cell types.
Purity
>98%
Description
Green fluorescent membrane permeable calcium indicator
Figure 1. ATP induced Ca2+ release in CHO-K1 cells measured using Fluo-4 AM
ATP causes a dose dependent release of Ca2+ from CHO K1 cells which is detected as an increase in fluorescence from Fluo-4. Method. CHO K1 cells were cultured and loaded with Fluo-4 AM following standard protocols (see our Fluo-4 AM Protocol). After a baseline, differing concentrations of ATP were added to cells at 30 seconds before the change in fluorescence was measured for a subsequent 90 seconds.
Figure 2. ΔF following ATP application to CHO-K1 cells.
Representative image showing the increase in Fluo-4 mediated fluorescence following incubation of CHO-K1 cells with ATP.
Figure 1. ATP induced Ca2+ release in CHO-K1 cells measured using Fluo-4 AM
ATP causes a dose dependent release of Ca2+ from CHO K1 cells which is detected as an increase in fluorescence from Fluo-4. Method. CHO K1 cells were cultured and loaded with Fluo-4 AM following standard protocols (see our Fluo-4 AM Protocol). After a baseline, differing concentrations of ATP were added to cells at 30 seconds before the change in fluorescence was measured for a subsequent 90 seconds.
Figure 2. ΔF following ATP application to CHO-K1 cells.
Representative image showing the increase in Fluo-4 mediated fluorescence following incubation of CHO-K1 cells with ATP.
This compound is light sensitive; exposure to light may affect compound performance. We therefore recommend storing the solid material and any solutions in the dark and protecting from light.
Important
This product is for RESEARCH USE ONLY and is not intended for therapeutic or diagnostic use. Not for human or veterinary use
Can I fix my cells after loading with Fluo-4 AM dye for Ca2+ detection?
No. Fluo-4 AM isn't covalently bound to any cellular components and fixation damages the membrane meaning that the dye would leak out of the cell.
How can I prevent Fluo-4 leaking out of cells?
Fluo-4 AM generally has very good cell retention with minimal leakage. If excessive leakage is observed try either reducing the incubation temperature or adding probenecid to the loading solution.
Can unused dye be frozen and reused?
Once dye has been dissolved in DMSO then it is possible to aliquot and freeze at -20°C. Try to avoid more than 2 freeze/thaw cycles.
Fluo-4 AM Protocol (222.8 KB) 
