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Cell Meter™ No Wash and Probenecid-Free Endpoint Calcium Assay Kit
Optimized for microplate reader
Cell Meter™ No Wash and Probenecid-Free Endpoint Calcium Assay Kit enables homogeneous fluorescence-based assays for detecting intracellular calcium mobilization without the need to use kinetics reading mode. It can be used on Fluorescence microplate readers with bottom read mode that do not have a build-in liquid dispenser or the ability of kinetics reading. After loading the Fluo-8E™AM dye into cells of interest, without wash steps, one can simply add the calcium flux agonist by a liquid dispenser or hand pipetting, and then read the plate by a fluorescent reader. Fluo-8E™ AM can cross cell membrane passively by diffusion. Once inside the cells, the lipophilic blocking groups of Fluo-8E™ AM are cleaved by esterase, resulting in a negatively charged fluorescent dye that stays inside cells. Its fluorescence is greatly enhanced and long lasting upon binding to calcium. When cells expressing GPCR of interest are stimulated with an agonist, the receptor signals the release of intracellular calcium, which significantly increases the fluorescence of Fluo-8E™. The characteristics of its high sensitivity, >100 times fluorescence enhancement and long lasting fluorescent signal make Fluo-8E™ an ideal indicator for the measurement of cellular calcium on fluorescence microplate readers that do not have the fluid transfer and kinetic reading mode capability. The Cell Meter™ No Wash and Probenecid-Free Endpoint Calcium Assay Kit can be performed in a 96-well or 384-well microtiter-plate format.
The ATP dose dependent intracellular calcium release was measured by Cell Meter™ No Wash and Probenecid-Free Endpoint Calcium Assay Kit in CHO-K1 cells in a 96-well plate. 3 columns of cells were incubated with Fluo-8E™ AM dye loading solution for 1 hr at 37°C before ATP was added into all 3 columns of the wells. The plate was read immediately after the addition of the ATP by ClarioStar (BMG Labtech) at Ex/Em = 490/525 nm with bottom and endpoint read mode.
The ATP dose dependent intracellular calcium release was measured by Cell Meter™ No Wash and Probenecid-Free Endpoint Calcium Assay Kit in CHO-K1 cells in a 96-well plate. 3 columns of cells were incubated with Fluo-8E™ AM dye loading solution for 1 hr at 37°C before ATP was added into all 3 columns of the wells. The plate was read immediately after the addition of the ATP by ClarioStar (BMG Labtech) at Ex/Em = 490/525 nm with bottom and endpoint read mode.
protocol
Protocol
sds
SDS
COA
CatalogSize
Price
Quantity
36312100 Tests
Price
 
Spectral properties
Correction factor (260 nm)1.076
Correction factor (280 nm)0.769
Extinction coefficient (cm -1 M -1)
23430
Excitation (nm)495
Emission (nm)516
Quantum yield
0.16
1
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12352200
Instrument settings
Fluorescence microplate reader
Excitation490 nm
Emission525 nm
Cutoff515 nm
Recommended plateBlack wall/Clear bottom
Instrument specification(s)Bottom read mode
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Page updated on October 9, 2025