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 ℃) 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 ℃ 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.

1096.94

Solid

C₅₁H₅₀F₂N₂O₂₃

273221-67-3

Room temperature in continental US; may vary elsewhere.

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

*该产品在溶液状态不稳定，建议您现用现配，即刻使用。

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  Description: Fluo-4 AM can be used to determine total intracellular Ca²⁺levels.
  Method: For cell labeling.
  1. Count cells and take 10⁶cells from each sample.
  2. Collect cells (5 min, 3000×g, 4℃) and wash once in PBS.
  3. Resuspend cells in 0.5 mL PBS and add Fluo-4 AM (1 mM; 0.5 μL; 37℃; 1 h) to a final concentration of 1 μM.
  4. Wash cells three times (2 min, 3000×g) with PBS and finally resuspend in 1 mL PBS, and separated into 2 flow cytometry tubes-0.5 mL in each.
  5. Evaluate staining by flow cytometry and analyze the data by a software such as CellQuest software.