HERSTELLER

Calcium Assays


Classical Calcium Assay Reagents


Find AAT Bioquest reagents for assays of intracellular calcium at good prices here at www.biomol.de:


Ca2+ Detection by Green Fluorescence


Quest™ Fluo-8 Calcium Assay Kit for Cellular Analysis: Ideal as calcium assay, brighter than Fluo-3 and Fluo-4.


Ca2+ Detection by Red Fluorescence


Rhod-4: The Brightest and Most Sensitive Red Fluorescent Ca2+ Indicator: Large assay window - 10 times better than Rhod-2 AM for HTS applications.


New: Robust Homogeneous Fluorescence-based Calcium Detection with Cal-520™


Cal-520™ enables probenecid-free Ca2+ assays with significantly higher S/N ratio than any other commercially available fluorescent Ca2+ assays.


 


Comparison of Fluo-3 AM, Fluo-4 AM and Cal-520 AM 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(TM) calcium assay a robust tool for evaluating GPCR and calcium channel targets as well as for screening their agonists and antagonists. Cal 520™ AM technology has the following benefits and features:


Find more information in this poster.


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Quest™ Fluo-8 Calcium Assay Kit for Cellular Analysis


Ideal as calcium assay, brighter than Fluo-3 and Fluo-4.


 


Quest™ Fluo-8 Calcium Assay Kit for Cellular Analysis Utilizing the brightest and most sensitive fluorescent Ca2+ indicator, Screen Quest™ Fluo-8 Calcium Assay Kit provides much higher signal intensity and very large fluorescence intensity increase in response to Ca2+ binding. AAT Bioquest’s homogeneous fluorescence-based assay kit detects calcium mobilization across a broad spectrum of biological targets. Easy-to-use protocol does not require a wash step or exogenous addition of a quencher dye, which could adversely affect receptor-ligand interaction kinetics. Other widely used calcium ion indicators, such as Fluo-3, Fluo-4 and Calcium 4 dyes, have relatively weak fluorescence signals limiting their application in high-throughput screening applications. Quest Fluo-8™ dye offers superior measurement of challenging cell lines and receptors.





Features


Benefits



Load dye in live cells at room temperature (or 37°C)


Room temperature is more compatible with HTS applications


Homogeneous mix-and-read, no-wash calcium mobilization assay



Washing cells introduces errors



Significantly brighter fluorescence intensity


2X brighter than Fluo-4, Fluo-4 Direct, Fluo-4 NW, FLIPR® Calcium 4 assay kits
4X brighter than Fluo-3



EC50 values (half maximal effective concentration)


Comparable with Fluo-3, Fluo-4, Fluo-4 Direct™ and FLIPR® Calcium assay kits



No quencher dye


No potential interference from compound ligand-receptor interactions



Large assay window compared to other dyes


Easily detects low calcium mobilization in challenging cell lines and receptors


Maximum excitation at ~490 nm


Readily adopted into established protocols and instrument workflows


Rigorous Quality Control


Controls and eliminates non-specific assay artifacts



Summary


Compared to other commercial calcium assay kits based on either Fluo-3 or Fluo-4, AAT Bioquest\'s Screen Quest™ Calcium Assay Kits have the following advantages for HTS applications:


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Protocols:


Rhod-4: The Brightest and Most Sensitive Red Fluorescent Ca2+ Indicator


 


Rhod-4: The Brightest and Most Sensitive Red Fluorescent Ca2+ Indicator Since the 1920s, scientists have attempted to measure Ca2+, but few were successful due to limited availability of Ca2+ probes. The first reliable measurements of Ca2+ were performed by Ridgway and Ashley by injecting the photoprotein aequorin into the giant muscle fiber of the barnacle. Subsequently, in the 1980s, Tsien and colleagues produced a variety of fluorescent indicators. Among them the rhodamine-based Ca2+ reagents (such as Rhod-2) have been the most valuable longer wavelength dye for measuring Ca2+ with red fluorescence. Although Rhod-2 has been the most popular red fluorescent Ca2+ indicator, its mitochondrial localization and high basal Ca2+ signal in cells have severely limited its cellular applications. In addition, the less optimal excitation of Rhod-2 at 488 nm makes it less robust to use with some instruments (such as FLIPR™) that have only 488 nm excitation light source. Quest Rhod-4™ serial calcium detection reagents have been developed to address these limitations of Rhod-2. The absorption and emission peaks of Quest Rhod-4™ reagents are 530 nm and 555 nm, respectively. Although Rhod-4 has maximum absorption at 530 nm, its absorption at 488 nm is quite strong. It is quite unique that Quest Rhod-4™ can be well excited with an argon ion laser at 488 nm besides the longer wavelength 514 nm, 532 nm and 546 nm excitations. Quest Rhod-4™ emits fluorescence (at wavelengths 555 nm) that increases with increasing Ca2+. Quest Rhod-4™ is determined to undergo a >200-fold increase in fluorescence upon binding Ca2+. Because the range of increase in Ca2+ in many cells after stimulation is generally 5- to10-fold, Quest Rhod-4™ is an excellent probe to use with high sensitivity in this region. The Kd of Quest Rhod-4™ is estimated to be 525 nM (22°C, pH 7.0 –7.5), but this value may be significantly influenced by pH, viscosity, and binding proteins in vivo conditions.





Cat # Product Name Unit Size
ABD-21121 Quest Rhod-4™, AM 5x50 µg
ABD-21123 Quest Rhod-4™, AM 20x50 µg
ABD-21128 Quest Rhod-4™, sodium salt 5x50 µg
ABD-36331 Screen Quest™ Rhod-4 NW Calcium Assay Kit *Medium Removal* 10 Plates
ABD-36334 Screen Quest™ Rhod-4 NW Calcium Assay Kit *1% FBS Growth Medium * 10 Plates


Robust Homogeneous Fluorescence-based Calcium Detection with Cal-520™


 


Cal-520Download your product overview (pdf, 2 pages, 454 kB).


Reagents and Assay Kits for Calcium Functional Assays





Product Name
 BAPTA, AM
 BAPTA, AM *UltraPure Grade*
 BAPTA, tetrapotassium salt
 BAPTA, tetrasodium salt
 Coelenterazine *UltraPure grade*
 Coelenterazine *UltraPure grade*
 Coelenterazine cp *UltraPure grade*
 Coelenterazine cp *UltraPure grade*
 Coelenterazine f *UltraPure grade*
 Coelenterazine f *UltraPure grade*
 Coelenterazine h *UltraPure grade*
 Coelenterazine h *UltraPure grade*
 Coelenterazine h *UltraPure grade*
 Coelenterazine hcp *UltraPure grade*
 Coelenterazine hcp *UltraPure grade*
 Coelenterazine n *UltraPure grade*
 Coelenterazine n *UltraPure grade*
 Coelenterazine n *UltraPure grade*
 EGTA AM
 EGTA AM *10 mM DMSO solution*
 EGTA tetrasodium salt *10 mM aqueous solution*
 EGTA tetrasodium salt *UltraPure Grade*
 Fluo-3, AM
 Fluo-3, AM *Bulk package*
 Fluo-3, AM *Custom packaging*
 Fluo-3, AM *UltraPure grade*
 Fluo-3, pentaammonium salt
 Fluo-3, pentapotassium salt
 Fluo-3, pentasodium salt
 Fura-2, AM
 Fura-2, AM *Bulk packaging*
 Fura-2, AM *Custom packaging*
 Fura-2, AM *Custom packaging*
 Fura-2, AM *UltraPure Grade*
 Fura-2, pentapotassium salt
 Fura-2, pentasodium salt
 Indo-1, AM
 Indo-1, AM *Bulk packaging*
 Indo-1, AM *Custom packaging*
 Indo-1, AM *UltraPure Grade*
 Indo-1, pentapotassium salt
 Indo-1, pentasodium salt
 Phenol Red, sodium salt *Cell culture tested*
 Pluronic® F-127 *10% solution in water*
 Pluronic® F-127 *20% solution in DMSO*
 Pluronic® F-127 *Cell culture tested *
 Probenecid *Cell culture tested*
 Quest Fluo-8H™, AM
 Quest Fluo-8H™, AM
 Quest Fluo-8H™, AM
 Quest Fluo-8H™, sodium salt
 Quest Fluo-8L™, AM
 Quest Fluo-8L™, AM
 Quest Fluo-8L™, sodium salt
 Quest Fluo-8™, AM
 Quest Fluo-8™, AM
 Quest Fluo-8™, AM
 Quest Fluo-8™, AM
 Quest Fluo-8™, sodium salt
 Quest Rhod-4™, AM
 Quest Rhod-4™, AM
 Quest Rhod-4™, AM
 Quest Rhod-4™, AM
 Quest Rhod-4™, sodium salt
 Quest Rhod-4™, sodium salt
 Quin-2, AM
 Quin-2, tetrapotassium salt
 ReadiUse™ probenecid *25 mM stabilized aqueous solution*
 ReadiUse™ probenecid, sodium salt *Water-soluble*
 ReadiUse™ probenecid, sodium salt tablet *Water-soluble*
 Rhod-2, AM
 Rhod-2, AM *UltraPure Grade*
 Rhod-2, AM *UltraPure Grade* *Bulk packaging*
 Rhod-2, AM *UltraPure Grade* *Custom packaging*
 Rhod-2, tripotassium salt
 Rhod-2, trisodium salt
 Rhod-5N, AM
 Rhod-5N, tripotassium salt
 Screen Quest™ 10X calcium assay buffer with Phenol Red Plus™
 Screen Quest™ 10X cell staining buffer with Phenol Red Plus™
 Screen Quest™ 10X cell staining buffer with Phenol Red Plus™
 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 *Optimized for Difficult Cell Lines*
 Screen Quest™ Fluo-8 No Wash Calcium Assay Kit
 Screen Quest™ Fluo-8 No Wash Calcium Assay Kit
 Screen Quest™ Fluo-8 No Wash Calcium Assay Kit
 Screen Quest™ Luminometric Calcium Assay Kit
 Screen Quest™ Luminometric Calcium Assay Kit *10X10 Plates*
 Screen Quest™ Rhod-4 No Wash Calcium Assay Kit *1% FBS Growth Medium * *10 plates*
 Screen Quest™ Rhod-4 No Wash Calcium Assay Kit *1% FBS Growth Medium* *1 plate*
 Screen Quest™ Rhod-4 No Wash Calcium Assay Kit *1% FBS Growth Medium* *10x10 plates*
 Screen Quest™ Rhod-4 No Wash Calcium Assay Kit *Medium Removal* *1 plate*
 Screen Quest™ Rhod-4 No Wash Calcium Assay Kit *Medium Removal* *10 plates*
 Screen Quest™ Rhod-4 No Wash Calcium Assay Kit *Medium Removal* *10x10 plates*
 Trypan Blue, sodium salt *Cell culture tested*
 Trypan Blue, sodium salt *Cell culture tested*
 Trypan Blue, sodium salt *UltraPure grade* *Purified to eliminate fluorescent impurities*
 Trypan Purple™ *0.1 M aqueous solution*
 Trypan Purple™ *0.1 M aqueous solution*
 Trypan Red Plus™ *0.1 M aqueous solution*
 Trypan Red Plus™ *0.1 M aqueous solution*
 Trypan UltraBlue™, sodium salt *0.1 M aqueous solution*
 Trypan UltraRed™ *0.1 M aqueous solution*
 Trypan UltraRed™ *0.1 M aqueous solution*

Pluronic is a registered trademark of BASF SE. Download your Calcium Detection Reagents brochure (pdf, 130 kB).


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