Screen Quest™ Fluo-8® Medium Removal Calcium Assay Kit *Optimized for Difficult Cell Lines*

Screen Quest™ Fluo-8® Medium Removal Calcium Assay Kit

Optimized for Difficult Cell Lines

Screen Quest™ Fluo-8® Medium Removal Calcium Assay Kit is a fluorescence-based kit for monitoring intracellular calcium mobilization in cell-based microplate assays.

Calcium-responsive indicator: Fluo-8® NW fluorescence increases upon calcium binding following intracellular esterase cleavage of its cell-permeable form
Medium-removal format: Designed to reduce background fluorescence and interference associated with serum or culture medium
Microplate compatibility: Supports 96-well and 384-well formats and automated fluorescence detection
Research applications: Suitable for screening calcium-coupled GPCRs and evaluating calcium channel–associated responses

Carbachol Dose Response was measured in HEK-293 cells with Screen Quest™ Fluo-8 NW Assay Kit and Fluo-4 NW Assay Kit. HEK-293 cells were seeded overnight at 40,000 cells/100 µL/well in a Costar black wall/clear bottom 96-well plate. The growth medium was removed, and the cells were incubated with 100 µL of dye-loading solution using the Screen Quest™ Fluo 8-NW calcium assay kit or the Fluo-4 NW kit (according to the manufacturer's instructions) for 1 hour at room temperature. Carbachol (25µL/well) was added by NOVOstar (BMG Labtech) to achieve the final indicated concentrations. The EC50 of Carbachol using Fluo8 NW is about 1.2 µM.

Protocol

Catalog	Size	Price
36307	1 Plate	Price
36308	10 Plates	Price
36309	100 Plates	Price

Overview

Screen Quest™ Fluo-8® Medium Removal Calcium Assay Kit detects changes in intracellular calcium through the calcium-responsive fluorescence of Fluo-8® NW. The cell-permeable indicator enters cells, where intracellular esterases cleave its lipophilic blocking groups to generate a charged calcium indicator. Calcium binding increases fluorescence, providing a readout of cellular calcium responses.

The medium-removal format is designed to reduce background fluorescence and interference associated with serum or culture medium. The assay supports 96-well and 384-well microplates and automated fluorescence detection, with recommended reader settings of Ex/Em = 490/525 nm. Assay conditions and compound responses require empirical optimization for the selected cell model. The kit is suitable for high-throughput screening of calcium-coupled GPCRs and functional studies of calcium channels.

Components

Catalog 36307 (1 Plate)

Component A: Fluo-8® NW	1 vial, lyophilized
Component B: 10X Pluronic® F127 Plus	1 bottle (1 mL)
Component C: HHBS (Hanks' buffer with 20 mM Hepes)	1 bottle (9 mL)

Catalog 36308 (10 Plates)

Component A: Fluo-8® NW	1 vial, lyophilized
Component B: 10X Pluronic® F127 Plus	10 bottles (1 mL/bottle)
Component C: HHBS	1 bottle (100 mL)

Catalog 36309 (100 Plates)

Component A: Fluo-8® NW	10 vials, lyophilized
Component B: 10X Pluronic® F127 Plus	10 bottles (10 mL/bottle)
Component C: HHBS	Not included

Citations

View all 163 citations: Citation Explorer

COVID-19-associated neuroinflammation and astrocyte death in the brain linked to ORF3a-induced activation of Sur1-mediated ion channels

Authors:

Gerzanich, Volodymyr and Zhang, Chenyu and Zhang, Jiantao and Sallapalli, Bhargava Teja and Pei, Shaokai and Nasr, Mohamed and Tosun, Cigdem and Zhang, Yanjin and Tang, Qiyi and Simard, J Marc and others,

Journal:

mBio (2025): e02012--25

Piezo1 promotes the progression of necrotizing enterocolitis by activating the Ca2 (+)/CaMKII-dependent pathway

Authors:

Li, Zhenyu and Jiang, Qinlei and Wei, Jiaqi and Dang, Dan and Meng, Zhaoli and Wu, Hui

Journal:

Communications Biology (2025): 417

Transient receptor potential vanilloid 1 interacts with TLR4/CD14 signaling pathway in lipopolysaccharide-mediated inflammation in macrophages

Authors:

Hsu, Julia Chu-Ning and Tseng, Hsu-Wen and Chen, Chia-Hui and Lee, Tzong-Shyuan

Journal:

Experimental Animals (2024): 23--0148

Structural basis of antibody inhibition and chemokine activation of the human CC chemokine receptor 8

Authors:

Sun, Dawei and Sun, Yonglian and Janezic, Eric and Zhou, Tricia and Johnson, Matthew and Azumaya, Caleigh and Noreng, Sigrid and Chiu, Cecilia and Seki, Akiko and Arenzana, Teresita L and others,

Journal:

Nature Communications (2023): 7940

The potent analgesia of intrathecal 2R, 6R-HNK via TRPA1 inhibition in LF-PENS-induced chronic primary pain model

Authors:

Liu, An-Ran and Lin, Zhen-Jia and Wei, Ming and Tang, Yuan and Zhang, Hui and Peng, Xiang-Ge and Li, Ying and Zheng, Yu-Fan and Tan, Zhi and Zhou, Li-Jun and others,

Journal:

The Journal of Headache and Pain (2023): 1--27

References

View all 26 references: Citation Explorer

Novel fluo-4 analogs for fluorescent calcium measurements

Authors:

Martin VV, Beierlein M, Morgan JL, Rothe A, Gee KR.

Journal:

Cell Calcium (2004): 509

Kinetic characterization of novel NR2B antagonists using fluorescence detection of calcium flux

Authors:

Bednar B, Cunningham ME, Kiss L, Cheng G, McCauley JA, Liverton NJ, Koblan KS.

Journal:

J Neurosci Methods (2004): 247

Flow cytometric kinetic assay of calcium mobilization in whole blood platelets using Fluo-3 and CD41

Authors:

do Ceu Monteiro M, Sansonetty F, Goncalves MJ, O'Connor JE.

Journal:

Cytometry (1999): 302

Amplitude distribution of calcium sparks in confocal images: theory and studies with an automatic detection method

Authors:

Cheng H, Song LS, Shirokova N, Gonzalez A, Lakatta EG, Rios E, Stern MD.

Journal:

Biophys J (1999): 606

A simple numerical model of calcium spark formation and detection in cardiac myocytes

Authors:

Smith GD, Keizer JE, Stern MD, Lederer WJ, Cheng H.

Journal:

Biophys J (1998): 15

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

Usage and storage

Intended use

Research Use Only (RUO)

Instrument settings

Fluorescence microplate reader
Excitation	490 nm
Emission	525 nm
Cutoff	510 nm
Recommended plate	Black wall/clear bottom
Instrument specification(s)	Bottom read mode/Programmable liquid handling

Other instruments	ArrayScan, FDSS, FLIPR, FlexStation, IN Cell Analyzer, NOVOStar, ViewLux

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 7, 2026