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Cal-520ER™ AM

Cal-520ER™ has been designed to monitor the change of calcium ion (Ca2+) in the endoplasmic reticulum (ER). Cal-520ER™ AM is cell permeable, thus can be readily used in live cells. The importance of calcium signaling in cell health and disease is the major driving force in current research of intracellular calcium homeostasis. Ca2+ release from ER and other calcium stores seems to be the crucial factor in the activation of many cellular functions. Significant changes in ER Ca2+ content and dynamics have been implicated in the activation of the ER stress response, abnormal autophagy, and cell death which leads to a variety of pathological conditions. Cal-520ER is a low-affinity Ca2+ indicator that can be used to record fast Ca2+ signals and to measure the kinetics of Ca2+ currents. Compared to Oregon Green BAPTA-5N and to Fluo4FF, Cal-520ER offers a superior signal-to-noise ratio providing the optimal characteristics for this important type of biophysical measurement. This ability is the result of a relatively high fluorescence at zero Ca2+, necessary to detect enough photons at short exposure windows, and a high dynamic range leading to large fluorescence transients associated with short Ca2+ influx periods.

Endogenous P2Y receptor response to ATP in CHO-K1 cells. CHO-K1 cells were seeded overnight at a density of 40,000 cells per 100 µL per well in a 96-well black wall/clear bottom Costar plate. Next, 100 µL of 10 µM Cal-520ER™ AM in HHBS with 1 mM probenecid was added to each well. The cells were then incubated at 37 °C for 3 hours. In the final 30 minutes of incubation, the Cell Navigator® Live Cell Endoplasmic Reticulum (ER) Staining Kit *Red Fluorescence* (Cat# 22636) was added to stain the ER. After incubation, the dye-loading mediums were replaced with 100 µL HHBS containing 1 mM probenecid. The cells were imaged with a fluorescence microscope (Olympus IX71) using the FITC channel, both before and after the addition of 50 µL of 300 µM ATP.
Endogenous P2Y receptor response to ATP in CHO-K1 cells. CHO-K1 cells were seeded overnight at a density of 40,000 cells per 100 µL per well in a 96-well black wall/clear bottom Costar plate. Next, 100 µL of 10 µM Cal-520ER™ AM in HHBS with 1 mM probenecid was added to each well. The cells were then incubated at 37 °C for 3 hours. In the final 30 minutes of incubation, the Cell Navigator® Live Cell Endoplasmic Reticulum (ER) Staining Kit *Red Fluorescence* (Cat# 22636) was added to stain the ER. After incubation, the dye-loading mediums were replaced with 100 µL HHBS containing 1 mM probenecid. The cells were imaged with a fluorescence microscope (Olympus IX71) using the FITC channel, both before and after the addition of 50 µL of 300 µM ATP.
protocol
Protocol
sds
SDS
COA
CatalogSize
Price
Quantity
2114910x50 ug
Price
 
Physical properties
Molecular weight1183.94
SolventDMSO
Spectral properties
Excitation (nm)492
Emission (nm)515
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
Instrument settings
Fluorescence microscope
ExcitationFITC
EmissionFITC
Recommended plateBlack wall/clear bottom
Fluorescence microplate reader
Excitation490
Emission525
Cutoff515
Recommended plateBlack wall/clear bottom
Instrument specification(s)Bottom read mode/Programmable liquid handling
Documents
Protocol
Safety Data Sheet
Certificate of Analysis
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Page updated on October 18, 2025