Description
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 Bright is a cell permeable Ca2+ indicator which is metabolized by intracellular esterase and results in a negatively charged fluorescent dye. PhenoVue Cal-520 AM Bright corresponds to an improved version of the PhenoVue Cal-520 AM which exhibits higher brightness and signal-to-noise, higher intracellular retention and may not require probenecid, compared to other green calcium indicators such as Fluo-4 AM or PhenoVue Cal-520 AM. PhenoVue Cal-520 AM Bright is the most robust Ca2+ indicator.
Typical working concentration: 5 µM
Equivalent number of microplates:
3-10 x 96-well microplates
3-10 x 384-well microplates
4-16 x 1536-well microplates
