1. Dye Reagents
  2. Fluo-4 AM

Fluo-4 AM 

Cat. No.: HY-101896
Handling Instructions

Fluo-4 AM is a cell-permeable Ca2+ indicator.

For research use only. We do not sell to patients.

Fluo-4 AM Chemical Structure

Fluo-4 AM Chemical Structure

CAS No. : 273221-67-3

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  • Biological Activity

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Fluo-4 AM is a cell-permeable Ca2+ indicator[1].

In Vitro

Fluo-4 AM is a fluorescent dye (λex=494 nm, λem=516 nm). Preloaded with Fuo-4 AM, a very bright fluorescence image is observed. In a parallel experiment with fluo-3 AM-loaded cells, the resulting fluorescence image, although clearly discernable in this case, is less bright[1]. Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
1. Count the cells and take 106 cells from each sample (control and experiment/s).
2. Collect the cells (5 min, 3000×g, 4 °C) and wash once in PBS.
3. Resuspend the cells in 0.5-ml PBS and add 0.5 μl of Fluo-4-AM (1 mM stock) to a final concentration of 1 μM. Incubate at 37 °C for 1 h.
4. Wash the cells three times (2 min, 3000×g) with PBS and finally resuspend in 1-ml PBS. Separate into 2 flow cytometry tubes—0.5 ml in each.
5. Evaluate the staining by flow cytometry and analyze the data by a software such as CellQuest software.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Molecular Weight





Room temperature in continental US; may vary elsewhere.


-20°C, sealed storage, away from moisture and light

*The compound is unstable in solutions, freshly prepared is recommended.

Cell Assay

For measuring fluorescence from cells in suspension, dilutions to 2 to 3×106 cells are made, from cultures of rat basophilic leukemia (RBL) cells. Cells are incubated in suspension in 1 μM dye (including Fluo-4 AM) for 30 min at 37°C. Cell suspensions are then transferred to cuvets for measurements of fluorescence emission intensity by spectrofluorometer[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.


Purity: ≥99.0%

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