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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Description

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

1096.94

Formula

C51H50F2N2O23

CAS No.
Shipping

Room temperature in continental US; may vary elsewhere.

Storage

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

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

References
Cell Assay
[1]

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.

References

Purity: ≥99.0%

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