Cal-520FF™, AM

Cal-520® provides a robust homogeneous fluorescence-based assay tool for detecting intracellular calcium mobilization. Cal-520® AM is a new fluorogenic calcium-sensitive dye with a significantly improved signal to noise ratio and intracellular retention compared to the existing green calcium indicators (such as Fluo-3 AM and Fluo-4 AM). Cells expressing a GPCR or calcium channel of interest that signals through calcium can be preloaded with Cal-520® AM which can cross cell membrane. Once inside the cell, the lipophilic blocking groups of Cal-520™AM are cleaved by esterases, resulting in a negatively charged fluorescent dye that stays inside cells. Its fluorescence is greatly enhanced upon binding to calcium. When cells stimulated with agonists, the receptor signals the release of intracellular calcium, which significantly increase the fluorescence of Cal-520®. The characteristics of its long wavelength, high sensitivity, and >100 times fluorescence enhancement, make Cal-520® AM an ideal indicator for the measurement of cellular calcium. The high S/N ratio and better intracellular retention make the Cal-520® calcium assay a robust tool for evaluating GPCR and calcium channel targets as well as for screening their agonists and antagonists. Compared to other Cal-520® indicators, Cal-520FF™ has the lowest affinity to calcium with Kd ~ 10 uM.

Protocol

COA

Catalog	Size	Price	Quantity
21142	1 mg	Price
21143	10x50 ug	Price

Calculators

Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of Cal-520FF™, AM to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

	0.1 mg	0.5 mg	1 mg	5 mg	10 mg
1 mM	87.802 µL	439.012 µL	878.025 µL	4.39 mL	8.78 mL
5 mM	17.56 µL	87.802 µL	175.605 µL	878.025 µL	1.756 mL
10 mM	8.78 µL	43.901 µL	87.802 µL	439.012 µL	878.025 µL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
mg	÷	1138.92 g/mol	=	mL	x	mM	=	mmol

Citations

View all 95 citations: Citation Explorer

KS0365, a novel activator of the transient receptor potential vanilloid 3 (TRPV3) channel, accelerates keratinocyte migration

Authors:

Maier, Marion and Olthoff, Stefan and Hill, Kerstin and Zosel, Carolin and Magauer, Thomas and Wein, Lukas Anton and Schaefer, Michael

Journal:

British Journal of Pharmacology (2022): 5290--5304

Cal-520FF is the Present Optimal Ca 2+ Indicator for Ultrafast Ca 2+ Imaging and Optical Measurement of Ca 2+ Currents

Authors:

Bl{\"o}mer, Laila Ananda and Filipis, Luiza and Canepari, Marco

Journal:

Journal of Fluorescence (2021): 619--623

Synergistic drug combination effectively blocks Ebola virus infection

Authors:

Sun, Wei and He, Shihua and Martínez-Romero, Carles and Kouznetsova, Jennifer and Tawa, Gregory and Xu, Miao and Shinn, Paul and Fisher, Ethan G and Long, Yan and Motabar, Omid and others, undefined

Journal:

Antiviral Research (2017): 165--172

High-Throughput Phenotyping of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes and Neurons Using Electric Field Stimulation and High-Speed Fluorescence Imaging

Authors:

Daily, Neil J and Du, Zhong-Wei and Wakatsuki, Tetsuro

Journal:

ASSAY and Drug Development Technologies (2017)

HTS-Compatible Voltage-and Ca 2+-Sensitive Dye Recordings from hiPSC-Derived Cardiomyocytes Using the Hamamatsu FDSS Systems

Authors:

Kettenhofen, Ralf

Journal:

Stem Cell-Derived Models in Toxicology (2017): 135--152

Physical properties

Dissociation constant (K_d, nM)	9800
Molecular weight	1138.92
Solvent	DMSO

Spectral properties

Excitation (nm)	492
Emission (nm)	515
Quantum yield	0.75 ¹

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	12352200

Instrument settings

Fluorescence microscope
Excitation	FITC
Emission	FITC
Recommended plate	Black wall/clear bottom

Fluorescence microplate reader
Excitation	490
Emission	525
Cutoff	515
Recommended plate	Black wall/clear bottom
Instrument specification(s)	Bottom read mode/Programmable liquid handling

Telephone
Fax
Email	sales@aatbio.com
International	See distributors
Bulk request	Inquire
Custom size	Inquire
Technical Support	Contact us
Request quotation	Request
Purchase order	Send to sales@aatbio.com
Shipping	Standard overnight for United States, inquire for international

Page updated on October 6, 2025