Screen Quest™ Calbryte-520 Probenecid-Free and Wash-Free Calcium Assay Kit

Screen Quest™ Calbryte™ 520 Probenecid-Free and Wash-Free Calcium Assay Kit is a homogeneous fluorescence-based calcium assay kit for detecting intracellular calcium mobilization in live cells without probenecid or wash steps.

Probenecid-free workflow: Uses Calbryte™ 520 AM, a calcium indicator designed for improved intracellular retention without requiring probenecid
Wash-free assay format: Supports homogeneous calcium flux detection without dye-removal or washing steps
Calcium-responsive fluorescence: Calbryte™ 520 AM is converted by intracellular esterases into a calcium-sensitive dye with increased fluorescence upon calcium binding
Research applications: Suitable for GPCR calcium signaling, calcium channel analysis, difficult cell lines, and high-throughput screening in 96-well or 384-well formats

Comparison of fluorescent signal response of endogenous P2Y receptor to ATP in CHO-K1 cells. CHO-K1 cells were seeded overnight at 50,000 cells/100 µL/well in a 96-well black wall/clear bottom costar plate. Calcium flux response was measured with Screen Quest™ Calbryte™ 520 Probenecid-Free and Wash-Free Calcium Assay Kit, FLIPR Calcium 4 Assay Kit , and Fluo-4 Direct Calcium Assay kit. ATP (50 µL/well) was added by FlexStation 3 to achieve the final indicated concentrations.

Protocol

Catalog	Size	Price
36317	1 Plate	Price
36318	10 Plates	Price
36319	100 Plates	Price

Overview

Screen Quest™ Calbryte™ 520 Probenecid-Free and Wash-Free Calcium Assay Kit is designed for fluorescence-based detection of intracellular calcium mobilization in live cells. Calcium flux assays are widely used in drug discovery for screening G protein-coupled receptors (GPCRs) and calcium channels. The kit uses Calbryte™ 520 AM, a cell-permeant calcium indicator that enters cells and is converted by intracellular esterases into a calcium-responsive fluorescent dye.

The Calbryte™ 520 dye is designed for improved cellular retention, enabling calcium flux detection without probenecid and without wash steps. When cells are stimulated with screening compounds, receptor-mediated calcium release or calcium influx produces an increase in green fluorescence that can be measured in a microplate format. The homogeneous assay workflow is compatible with 96-well and 384-well formats and can be adapted to automated screening systems. Screen Quest™ Calbryte™ 520 Probenecid-Free and Wash-Free Calcium Assay Kit is suitable for GPCR calcium signaling studies, calcium channel assays, high-throughput screening, and calcium flux analysis in fragile or difficult cell lines.

Components

Catalog 36317 (1 Plate)

Component A: Calbryte™ 520 AM	1 vial
Component B: 10X Pluronic® F127 Plus	1 vial (1 mL)
Component C: HHBS (Hanks’ buffer with 20 mM Hepes)	1 vial (9 mL)

Catalog 36318 (10 Plates)

Component A: Calbryte™ 520 AM	1 vial
Component B: 10X Pluronic® F127 Plus	1 bottle (10 mL)
Component C: HHBS	1 bottle (100 mL)
Component C: HHBS (Hanks’ buffer with 20 mM Hepes)	1 bottle (100 mL)

Catalog 36319 (100 Plates)

Component A: Calbryte™ 520 AM	10 vials
Component B: 10X Pluronic® F127 Plus	10 bottles (10 mL/bottle)
Component C: HHBS	Not provided

Citations

View all 16 citations: Citation Explorer

OPEN ACCESS EDITED BY

Authors:

Taggart, Clifford and Kesturu, Girish S and Brown, Ryan and Wu, Xiaojun and Wang, Zhengtao and Chen, Z and Wang, G and Xie, X and Liu, H and Liao, J and others,

Journal:

Oral neutrophils-the good, the bad, and the ugly (2023): 41

Preferential Gs protein coupling of the galanin Gal1 receptor in the $\mu$-opioid-Gal1 receptor heterotetramer

Authors:

De Oliveira, Paulo A and Moreno, Estefan{\'\i}a and Casajuana-Martin, Nil and Casad{\'o}-Anguera, Ver{\`o}nica and Cai, Ning-Sheng and Camacho-Hernandez, Gisela Andrea and Zhu, Hu and Bonifazi, Alessandro and Hall, Matthew D and Weinshenker, David and others,

Journal:

Pharmacological Research (2022): 106322

Selective activation of TRPA1 ion channels by nitrobenzene skin sensitizers DNFB and DNCB

Authors:

Wu, Han and Niu, Canyang and Qu, Yaxuan and Sun, Xiaoying and Wang, KeWei

Journal:

Journal of Biological Chemistry (2022)

Selective activation of TRPA1 ion channel by skin sensitizer nitrobenzene DNFB

Authors:

Wu, Han and Qu, Yaxuan and Sun, Xiaoying and Wang, Kewei

Journal:

Authorea Preprints (2021)

Diquafosol tetrasodium elicits total cholesterol release from rabbit meibomian gland cells via P2Y 2 purinergic receptor signalling

Authors:

Endo, Ken-ichi and Sakamoto, Asuka and Fujisawa, Koushi

Journal:

Scientific reports (2021): 1--8

References

View all 172 references: Citation Explorer

Calcium imaging of cortical neurons using Fura-2 AM

Authors:

Barreto-Chang OL, Dolmetsch RE.

Journal:

J Vis Exp. (2009)

A flow cytometric comparison of Indo-1 to fluo-3 and Fura Red excited with low power lasers for detecting Ca(2+) flux

Authors:

Bailey S, Macardle PJ.

Journal:

J Immunol Methods (2006): 220

Measurement of [Ca2+]i in whole cell suspensions using fura-2

Authors:

Hirst RA, Harrison C, Hirota K, Lambert DG.

Journal:

Methods Mol Biol (2006): 37

Load of calcium probe Fura -2/AM in Escherichia coli cells

Authors:

Shao M, Wang HM, Liu ZH, Shen P, Cai RX.

Journal:

Wei Sheng Wu Xue Bao (2005): 805

Fluorescence measurements of free [Mg2+] by use of mag-fura 2 in Salmonella enterica

Authors:

Froschauer EM, Kolisek M, Dieterich F, Schweigel M, Schweyen RJ.

Journal:

FEMS Microbiol Lett (2004): 49

Spectral properties

Excitation (nm)	493
Emission (nm)	515
Quantum yield	0.75 ¹

Usage and storage

Intended use

Research Use Only (RUO)

Instrument settings

Fluorescence microplate reader
Excitation	490 nm
Emission	525 nm
Cutoff	515 nm
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 September 29, 2026