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  3. Fluo-4 AM

Fluo-4 AM 

Cat. No.: HY-101896 Purity: ≥98.0%
COA 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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100 μg USD 150 In-stock

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This product is a controlled substance and not for sale in your territory.

Customer Review

Based on 15 publication(s) in Google Scholar

Top Publications Citing Use of Products

    Fluo-4 AM purchased from MCE. Usage Cited in: Front Pharmacol. 2022 May 17;13:855626.  [Abstract]

    Intracellular Ca2+ is measured by Fluo-4/AM (for 30 min) with flow cytometry.

    Fluo-4 AM purchased from MCE. Usage Cited in: Cell Death Discov. 2021 Feb 10;7(1):31.  [Abstract]

    The cardiomyocytes are isolated by enzyme digestion and loaded with Fluo 4-AM (5 μM) at 37 °C for 20 min. Fluorescence changes of Fluo 4-AM are recorded by confocal microscopy at 25 °C.

    Fluo-4 AM purchased from MCE. Usage Cited in: Oxid Med Cell Longev. 2021 Feb 10;2021:8884922.

    NP cells are incubated with 5 μM Fura-4-AM for 30 min at 37°C in the dark. Intracellular Ca2+ levels measured using the specific Ca2+-sensitive fluorescent indicator Fura-4-AM and analyzed via fluorescence microscopy.
    • Biological Activity

    • Protocol

    • Purity & Documentation

    • References

    • Customer Review


    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






    CAS No.
    Emission (Em)


    Excitation (Ex)





    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.

    Purity & Documentation

    Purity: ≥99.0%

    Dyeing Example
    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.

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    Fluo-4 AM Related Classifications

    Help & FAQs
    • Do most proteins show cross-species activity?

      Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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    The molarity calculator equation

    Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

    Mass   Concentration   Volume   Molecular Weight *
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    The dilution calculator equation

    Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

    This equation is commonly abbreviated as: C1V1 = C2V2

    Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
    × = ×
    C1   V1   C2   V2

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