Fluo-4 AM is a?cell-permeable Ca²⁺?indicator. Storage: protect from light.

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. 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.

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

• . 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]

  . Fluo-4.