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PhenoVue Cal-520 AM, Calcium Indicator

PhenoVue Cal-520 AM is a cell permeable fluorescent Ca2+ indicator and is part of Revvity’s portfolio of cellular imaging reagents. PhenoVue Cal-520 AM Ca2+ indicator exhibits green fluorescence and is validated for use in imaging microscopy and high-content screening applications. It exhibits a maximum excitation wavelength at 493 nm and a maximum emission wavelength of 515 nm.

View our extensive validation data in the Product Information Sheet within the Resources tab below.

For research use only, not for use in diagnostic procedures.
Part Number: CP125201
Part Number: CP125201
  • Calcium ions are important signaling molecules mediating a broad spectrum of intracellular, intercellular as well as extracellular functions. Ca2+ plays critical physiological functions such as muscle contraction, neuronal excitability, cell migration and cell growth. Intracellular Ca2+ is stored in the endoplasmic reticulum, sarcoplasmic reticulum, and the mitochondria. Upon cell surface receptors stimulation like GPCRs, calcium is released from these internal stores and its intracellular concentration increases from 100 nM to approximately 1 μM.

    Intracellular Ca2+ responses have been widely investigated with fluorescent calcium probes which fluorescence signal increases upon Ca2+ binding.

    The first-generation of fluorescent calcium indicators, including Fura-2, Fluo-3 and Rhod-2, were developed, followed by a second-generation, such as Fluo-4, with improved properties especially brightness and signal-to-noise ratio. However, the presence of anion transporters expressed at the cell surface may impair the intracellular retention of the fluorescent calcium indicators which are cleared out from cells. Inhibitors of these transporters, such as Probenecid, are commonly used but suffer from limitations such as cytotoxicity, off-targets, or inconsistent calcium flux kinetics. Such limitations can be overcome with third generation of fluorescent calcium indicators including Cal-520, Cal-590 and their corresponding Cal-520, Cal-590 Bright versions which do not require probenecid.

    PhenoVue Fluorescent Calcium Indicators, encompassing Fura-2 AM, Fluo-4 AM, Cal-520 AM, Cal-590 AM, Cal-520 AM Bright and Cal-590 AM Bright, enable a myriad of applications as well as multiplexing capability to unravel intracellular calcium mediated functions.

    PhenoVue Cal-520 AM is a cell permeable Ca2+ indicator which is metabolized by intracellular esterases and results in a negatively charged fluorescent dye. Consequently, PhenoVue Cal-520 AM exhibits higher sensitivity, significant improved signal-to-noise ratio, reduced leakage and higher intracellular retention compared to other green calcium indicators such as Fura-2 AM and Fluo-4 AM.


    • Numbers of Vials Per Unit: 10
    • Quantity or Volume Per Vial: 50 µg (45 nmoles)
    • Form: Solid
    • Storage: -16 °C
    • Recommended Working Concentration: 5 µM
    • Maximum Excitation Wavelength: 493 nm
    • Maximum Emission Wavelength: 515 nm
    • Common Filter Set: FITC
    • Live Cell Staining: Yes
    • Fixed Cell Staining: No

    Equivalent Number of Microplates:

    • 3-10 x 96-well microplates
    • 3-10 x 384-well microplates
    • 4-16 x 1536-well microplates
  • Color
    High Content Screening
    Imaging Microscopy
    Detection Method
    Product Compatibility
    Live samples only
    Shipping Conditions
    Shipped in Dry Ice
    Individual Reagent
    Unit Size
    10 vials


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Application Note
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