Fluo-4 AM 

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

Fluo-4 AM is a fluorescent dye (λ_ex=494 nm, λ_em=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.

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

1096.94

C₅₁H₅₀F₂N₂O₂₃

273221-67-3

Solid

Brown to red

526

485

O=C(OCOC(C)=O)CN(C1=CC=C(C2=C3C=C(F)C(C=C3OC4=C2C=C(F)C(OCOC(C)=O)=C4)=O)C=C1OCCOC5=CC(C)=CC=C5N(CC(OCOC(C)=O)=O)CC(OCOC(C)=O)=O)CC(OCOC(C)=O)=O

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.

• [1]. Gee KR, et al. Chemical and physiological characterization of fluo-4 Ca(2+)-indicator dyes. Cell Calcium. 2000 Feb;27(2):97-106.  [Content Brief]

  [2]. Fluo-4.