ION Biosciences is focused on producing a new generation of enhanced fluorescent indicators and assays. We work directly with customers to provide pure, high quality products that deliver better research results.

We know that your research is important, because at ION we are scientists too. We strive to develop meaningful solutions to the problems that stand in the way of your research goals. We start by putting the highest quality products in your hands, work hard to create truly useful and innovative solutions, and apply all our expertise to help support and ensure your success.

We at ION value the important work that you do. We value collaboration – we believe that together we are more effective at overcoming the most daunting research challenges. We value trust and integrity – the bedrock of successful collaboration. We value inclusion – we reject social, racial and religious intolerance. We value innovation – a key to creating a better future. We value stewardship of the environment – we practice recycling, waste reduction, and utilize green processes whenever possible.


Products

We combine our best fluorescent indicators with other critical reagents, rigorously validated protocols, and detailed, easy-to-follow instructions to provide industry-leading assays that outperform the competition and accelerate your research.

We offer a collection of membrane permeable and impermeable ION indicators targeting Ca²⁺, Na⁺, K⁺, TI⁺ as well as other fluorescent dyes to meet your research needs. Our indicators have the lowest lot-to-lot variability so you can have confidence in your results.

When the need to develop a validated assay for the most informative, relevant, and actionable results arises, researchers turn to ION Biosciences.
Free up valuable resources by outsourcing your assay development needs with ION.

Featured Products

ION is focused on producing a new generation of enhanced fluorescent indicators and assays to empower your research. We offer the highest quality products and the best prices.

Calcium

Calcium indicators are molecules that exhibit an increase in fluorescence upon binding Ca²⁺.

HeLa cells loaded with Fluo-4 in the presence (+) or the absence (-) of 500 nM of the Ca²⁺  ionophore ionomycin.

Ca²⁺- sensitive fluorescent indicators

Extensive research, including seminal contributions by ION’s principal scientists, has afforded a host of excellent, cell-compatible Ca²⁺ sensors. These molecules are small, synthetic fluorochromes which incorporate a Ca²⁺-binding moiety with excellent selectivity for Ca²⁺ over other physiologically relevant divalent cations, e.g. Mg²⁺. Under conditions where Ca²⁺ is not bound, the fluorescence of the sensor is significantly quenched. When Ca²⁺ is bound, the quenching is relieved, and the fluorescence of the sensor dramatically increases. For some sensors, binding Ca²⁺ also results in a change in the spectral properties of the indicator. For instance, in the case of Fura-2 (below), binding of Ca²⁺ results in a shift of the indicator’s excitation spectrum from a peak at 380 nm in the Ca²⁺ free form to 340 nm in the Ca²⁺-bound form. These indicators are often used in “ratiometric” mode where the Ca²⁺ binding-dependent changes in spectral properties provide a means to control things like photo-bleaching of the indicator and variable optical path length through samples.

Calcium-sensitive fluorescent indicators are compatible with a wide variety of detectors including fluorescent microscopes, plate readers, flow cytometers, and fluorescent indicator-doped solid-state sensors. Over the last 30 years, Ca²⁺-sensitive indicators have found numerous uses ranging from high-spatial resolution imaging of synaptic activity to high-throughput measurements of G protein-coupled receptor (GPCR) activation-evoked increases in intracellular Ca²⁺ for drug discovery.

 

Selecting a Ca²⁺ – sensitive fluorescent indicator

Choosing the correct indicator is crucial to the success of an experiment. A number of factors play a role in the decision including the indicator’s wavelength, its Ca²⁺ affinity as well as its cellular loading, retention and localization. ION provides a variety of the most popular and useful fluorescent Ca²⁺ sensors and reagents to help facilitate and empower your research.

These are the most popular forms of fluorescent Ca²⁺-sensitive indicators. They possess a range of Ca²⁺ affinities and the broadest instrument compatibility. We offer indicators with spectral properties well matched to common fluorescence microscopes, plate readers and flow cytometers using the most widely available FITC-compatible optics (Fluo-2, Fluo-3 and Fluo-4) while excitation sources and filters for Texas Red are compatible with ICR-1.

Fluo-2 

(EX 490 nm/Em 515 nm, Kd 290 nM) Also known as Fluo-2 HA, Fluo-2 offers the same spectral properties as Fluo-4 combined with superior loading in many cell types as well as a slightly higher Ca²⁺ affinity. The improved loading and increased affinity can result in superior sensitivity.

Fluo-3 

(EX 505 nm/Em 525 nm, Kd 390 nM) is a structurally related predecessor of Fluo-4 which has slightly lower affinity for Ca²⁺ and a slightly red-shifted excitation and emission spectrum. For nearly 10 years it was the most commonly used single wavelength, Ca²⁺ sensor. In most cell types Fluo-4 and Fluo-2 show superior loading compared to Fluo-3, however, some have reported that Fluo-3 is superior in certain instances.

Fluo-4 

(EX 490 nm/Em 515 nm, Kd 355 nM) is the most used and the most trusted of the single-wavelength Ca²⁺ indicators. It has been the indicator of choice for countless basic research studies and has been used to screen many tens-of-millions of compounds using high-throughput screening (HTS) to discover pharmacological tools and drugs for GPCRs, and a wide variety of Ca²⁺ permeable ion channels and transporters.

ICR-I

(EX 580 nm/Em 660 nm, Kd 480 nM) represents a new generation of long sought after fluorescent Ca²⁺ indicators that emit in the red portion of the spectrum. Because of ICR-1’s unique true-red spectral properties, it provides deeper tissue penetration and dramatic background reduction in tissues that have high autofluorescence, in the presence of serum or fluorescent compounds. ICR-1 is also particularly well-suited for use with the most common fluorescent proteins (e.g. GFP/YFP) as well as many optogenetic tools. Unlike some previous red-emitting Ca²⁺-sensitive indicators (e.g. Rhod-2) that concentrate in the mitochondria, ICR-1 enjoys a predominantly cytoplasmic localization similar to the Fluo family of indicators, making it ideal for most intracellular Ca²⁺ measurements.

These indicators are particularly useful when more accurate qualification of Ca²⁺ concentrations is desired. However, to utilize the benefits of ratiometric imaging, an instrument (e.g. fluorescence microscope) capable of rapid wavelength switching is required. In the case of Fura-2 (below) an excitation source and optics compatible with 340 nm and 380 nm illumination and ultraviolet (UV) light-compatible sample containers (e.g. microscope slides) are required.

Fura-2

(Ex 340/380 nm/505 nm, Kd 145 nM) is by far the mostly commonly used dual-wavelength excitation Ca²⁺-sensitive fluorescent indicator. While Fura-2’s affinity for Ca²⁺ is higher than any of the single wavelength indicators described above, it is still well-suited to measuring changes in cytoplasmic Ca²⁺ under most conditions while enjoying the benefits of ratiometry. In addition to traditional Fura-2, we also offer Fura-2 LR. Fura-2 LR is better retained in some cell lines compared to Fura-2 and thus, may provide superior results.

We provide our Ca²⁺-fluorescent dyes in a number of different forms and formats to best suit the intended application including membrane permeable forms, membrane impermeable forms and indicators packaged in kits together with other key reagents optimized to ensure best performance.

 

Cell permeable, acetoxymethyl group (AM) esters

 

When first introduced, loading the highly negatively charged Ca²⁺-sensitive fluorescent indicators was very challenging as they existed only as highly negatively charged, membrane impermeable salts. However, the development of forms of these molecules where the negative charges are masked by non-polar esters-linked moieties (AM esters) allow the molecules to enter cells through passive diffusion. An additional advantage of the AM form is that they are also non-fluorescent and non-Ca²⁺ binding. Once inside the cell, ubiquitous intracellular esterase enzymes promote rapid hydrolysis of the AM esters leaving the active, highly polar form of the indicator trapped inside the cell.

 

Reagents for improving indicator loading, retention and background suppression

 

Although the AM forms of ION’s fluorescent Ca²⁺ indicators readily cross the plasma membrane, there are other factors that can affect how well the indicators load into cells. Because AM forms of the indicators are quite hydrophobic, it is common to use the non-ionic surfactant, Pluronic F-127, to improve their solubility in aqueous solution thus improving their loading into cells. Pluronic F-127 at the concentrations employed for loading indicators into cells is generally well tolerated. While Pluronic F-127 isn’t absolutely essential for loading most fluorescent Ca²⁺ indicators into cells, it is highly recommended.

Another factor that can affect cell loading and intracellular localization is the presence of transport proteins capable of transporting many ion-sensitive, fluorescent indicators. In many cell types, the activity of these transporters can result in compartmentalization of the indicators into intracellular compartments as well as extrusion of the indicators outside of the cell. Both of these processes can dramatically reduce the signal-to-background levels and in some cell types (e.g. CHO cells),  make adequate loading of Ca²⁺-selective fluorescent indicators nearly impossible. Fortunately, inhibitors of these transport proteins can be an effective means of improving cytoplasmic localization and indicator retention. ION offers a convenient, 100X ION-Pro inhibitor solution that may be used in conjunction with any of our Ca²⁺-sensitive fluorescent indicators.

Calcium indicator kits and complete assay solutions

While fluorescent Ca²⁺-sensitive indicators have been in widespread use for decades, not every investigator is familiar with the most up-to-date protocols and additional reagents that may be necessary for their effective use. Therefore, ION provides customers with the option to purchase all reagents commonly used to load our Ca²⁺-sensitive fluorescent indicators into most cell types with clear, easy-to-follow instructions to facilitate successful use of fluorescent, Ca²⁺ sensitive indicators in one convenient package. These Brilliant kits are supplied in a variety of sizes ranging from trial packs to the largest-scale HTS (inquire).

Brilliant Calcium Express Kit

Our Express kit is custom-tailored to customer’s needs, providing only the reagents you need in the amounts you need them. Less waste, higher reproducibility, and more convenience.

Brilliant Calcium Express is a (EX 490 nm, EM 515 nm) High-throughput, no wash calcium (Ca²⁺) assay. The most flexible solution for measuring the activity of GPCRs coupled to intracellular Ca²⁺ release. 10 plates

Brilliant Calcium Flex Kit

Our Express kit is custom-tailored to customer’s needs, providing only the reagents you need in the amounts you need them. Less waste, higher reproducibility, and more convenience.

Brilliant Calcium Express is a (EX 490 nm, EM 515 nm) High-throughput, no wash calcium (Ca²⁺) assay. The most flexible solution for measuring the activity of GPCRs coupled to intracellular Ca²⁺ release. 10 plates

Ratiometric Calcium Essentials Kit

Ratiometric Calcium Essentials kit provides the necessary reagents for conducting no wash, ratiometric calcium flux assays compatible with plate reader and fluorescence microscopy applications. Individual components are provided to give the user the flexibility needed to customize your assay.

(EX 340/380 nm, EM 505 nm, Kd 145nM) High-throughput, no wash ratiometric calcium (Ca²⁺) assay. No wash, compatible with plate reader and fluorescence microscopy applications.

Be Brilliant! Explore our line of Calcium Indicators

Our collection of membrane permeable and impermeable ion indicators targeting Ca²⁺.

 

Product Description Ex/Em (nm) Kd (nM) Solubility Filter Set
BAPTA AM Non-fluorescent calcium (Ca²⁺) chelator, membrane permeable 250 DMSO
Brilliant Calcium Express High-throughput, no wash calcium (Ca²⁺) assay. An optimal solution for measuring intracellular Ca²⁺ dynamics, including effectors such as GPCRs and protein transporters. 10 plates 490/515 FITC, GFP
Brilliant Calcium Flex High-throughput, no wash calcium (Ca²⁺) assay. An optimal solution for measuring intracellular Ca²⁺ dynamics, including effectors such as GPCRs and protein transporters. 10 plates 490/515 FITC, GFP
Fluo-2 AM Green fluorescent, calcium (Ca²⁺) indicator, membrane permeable. Higher Ca²⁺ affinity than Fluo-4. 490/515 290 DMSO FITC, GFP
Fluo-2 K⁺ Salt Green fluorescent, calcium (Ca²⁺) indicator, membrane impermeable. Higher Ca²⁺ affinity than Fluo-4 490/515 290 H20 FITC
Fluo-3 AM Green fluorescent, calcium (Ca²⁺) indicator, membrane permeable 505/525 390 DMSO FITC
Fluo-3 K⁺ Salt Green fluorescent, calcium (Ca²⁺) indicator, membrane impermeable 505/525 390 H20 FITC
Fluo-4 AM Green fluorescent, calcium (Ca²⁺) indicator, membrane permeable 490/515 355 DMSO FITC, GFP
Fluo-4 K⁺ Salt Green fluorescent, calcium (Ca²⁺) indicator, membrane impermeable 490/515 355 H20 FITC
Fura-2 AM UV-excitable, ratiometric green calcium (Ca²⁺) indicator, membrane permeable 340,380/505 145 DMSO Fura-2
Fura-2 K⁺ Salt Ratiometric calcium (Ca²⁺) indicator, membrane impermeable 340,380/505 145 H20 Fura-2
Fura-2 LR AM Leakage-resistant, ratiometric calcium (Ca²⁺) indicator, membrane permeable 340,380/505 145 DMSO Fura-2
Fura-2 LR K⁺ Salt Leakage-resistant, ratiometric calcium (Ca²⁺) indicator, membrane impermeable 340,380/505 145 H20 Fura-2
ICR-1 AM Red fluorescent, calcium (Ca²⁺) indicator, membrane permeable. Optimal for multiplexing and tissue imaging. Fluorescence lifetime and multiphoton compatible. 580/660 480 DMSO Texas Red
ICR-1 K⁺ Salt Red fluorescent, calcium (Ca²⁺) indicator, membrane impermeable. Optimal for multiplexing and tissue imaging. Fluorescence lifetime and multiphoton compatible. 580/660 480 H20 Texas Red
ION Essentials Ratiometric Calcium The necessary reagents for conducting no wash, ratiometric calcium flux assays compatible with plate reader and fluorescence microscopy applications. Individual components are provided to give the user the flexibility needed to customize your assay. 340,380/505 145 DMSO GFP/FITC, PI/Texas Red

PH

BCECF AM

Ratiometric, green fluorescent pH indicator, membrane permeable.

SPECIFICATIONS

Excitation Emission Solubility:
430 490 nm 535 nm DMSO

 

Product Code:  Size: Product Description   Synonyms:
4011B 1mg BCECF AM 2′,7′-Bis-(2-Carboxyethyl)-5-(and-6)-Carboxyfluorescein – AM
4011E 20x5ug BCECF AM 2′,7′-Bis-(2-Carboxyethyl)-5-(and-6)-Carboxyfluorescein – AM

PRODUCT DESCRIPTION

BCECF AM is the most popular green fluorescent, intracellular pH indicator. BCECF has a pKa of ~7, and exhibits pH-dependent, dual-excitation properties (Ex/Em 430nm/535nm and 490nm/535nm) for ratiometric analysis. Ratiometry is optimal for imaging applications, and reduces effects of photobleaching, heterogenous dye loading, and variable cell morphology. For HTS applications, BCECF can also be used in non-ratiometric mode using standard fluorescein excitation and emission settings.

Potassium

Potassium indicators are molecules that exhibit an increase in fluorescence upon binding K⁺.

Potassium (K) in physiology

Potassium, along with sodium (Na⁺), is one of nature’s most important monovalent metal cations. Ion channels and transporters for K⁺ play a diverse range of functions from modulating neuronal activity to regulating solute balance. Potassium channels are by far the largest and most structurally diverse class of ion channels and much remains unknown about their functions in normal physiology and disease. An increasingly long list of K⁺ channels and K⁺ transporters are being associated with a wide variety of disorders.

Potassium channel and K⁺ transporter-targeted drugs already provide effective treatments for a wide breadth of indications including epilepsy, autism, type-2 diabetes, and diuresis. Unfortunately, K⁺ channels, particularly the Kv11.1 channel (a.k.a. hERG), may be the unintended target of other therapeutics resulting in sometimes deadly side-effects. As a result, pre-clinical testing of drug candidates to determine their effects on hERG channel activity is required.

Because expanding our knowledge of K⁺ channels and K⁺ transporters is of such intense interest to both basic and drug discovery researchers, a facile means to measure their activity is critical. Unfortunately, measuring intracellular and extracellular K⁺ levels and dynamics using fluorescent indicators is challenging. In most cells at rest, cytoplasmic K⁺ concentrations are above 100 mM while Na⁺ concentrations are in the 3 – 5 mM range. Conversely, extracellular K⁺ concentrations are typically in the 3-5 mM range while Na⁺ concentrations are above 100 mM. Therefore, an effective K⁺sensor needs to be able to discriminate between K⁺, Na⁺ and other common physiological cations while possessing a K⁺ affinity appropriate to detect physiologically relevant changes in K⁺ concentrations.

Extensive research has afforded the ION Potassium Green (IPG) family of excellent K⁺ sensors, formerly known as Asante Potassium Green (APG). These molecules are small, synthetic fluorochromes which incorporate a K⁺-binding moiety. Under conditions where K⁺ is not bound, the fluorescence of the sensor is significantly quenched. When K⁺ is bound, the quenching is relieved, and the fluorescence of the sensor dramatically increases.

The IPG family of indicators are the best-in-class replacements for the legacy K⁺ indicator, PBFI. Unlike PBFI, the spectral properties of the IPG family of K⁺ indicators make them convenient to use with common filter sets (e.g. YFP and FITC) as well as multiphoton approaches, and the range of K⁺ affinities make them useful for applications as diverse as extracellular K⁺ sensing to monitoring intracellular K⁺ dynamics. The IPG family of K⁺ indicators is compatible with a wide variety of detectors including fluorescent microscopes, plate readers, flow cytometers, and fluorescent indicator-doped solid-state sensors.

IPG-1

(EX 525 nm, EM 545 nm, Kd 50 mM) is the lowest affinity of the ION Potassium Green indicators. It is the indicator that is best positioned to respond to modest changes in intracellular K⁺ concentrations resulting from activation of plasma membrane K⁺ channels.

IPG-2

(EX 525 nm, EM 545 nm, Kd 18 mM) is the ION Potassium Green indicator with intermediate affinity. Currently, IPG-2 is our most popular intracellular K⁺ indicator.

IPG-4

(EX 525 nm, EM 545 nm, Kd 7 mM) has the highest K⁺ affinity of our single-wavelength indicators making it appropriate for conditions where there may be large changes in intracellular K⁺ (e.g. highly active synapses) or as an extracellular K⁺ sensor.

PBFI

(EX 340 nm, EM 380 nm, Kd 4 mM ) PBFI, AM is a potassium indicator that is ratiometric and UV light-excitable. This acetoxymethyl (AM) ester form is useful for noninvasive measurements of intracellular potassium levels.

While the IPG family of dyes are excellent fluorescent K⁺ indicators, the challenges of discriminating between K⁺ and Na⁺ are not trivial, and together with the often modest changes in intracellular or extracellular K⁺ results in relatively small signals compared to those observed with the Ca²⁺-sensitive fluorescent indicators, such as Fluo-4. In many instances, the Tl+ flux assay offers an outstanding alternative to K⁺-sensitive fluorescent indicators. The Tl⁺ flux assay takes advantage of a Tl⁺-selective fluorescent indicator, Thallos, and the fact that K⁺ channels and K⁺ transporters readily accept Tl⁺ as a K⁺ surrogate. When Tl⁺ is added to the outside of cells loaded with Thallos, Tl⁺ entering cells through K⁺ channels and K⁺ transporters results a dramatic increase in fluorescence. Today, the Tl⁺ assay is the most commonly used approach for large-scale high-throughput screening (HTS) of K⁺ channels and K⁺ transporters.

Thallos

(EX 490 nm, EM 515 nm) is the best-in-class Tl⁺-sensitive fluorescent indicator. It offers outstanding cell loading and excellent compatibility with commonly available FITC filters.

We provide our IPG family of K⁺-sensitive fluorescent indicators in membrane permeable and membrane impermeable forms. We offer Thallos as part of our convenient Brilliant Thallium Assay kits and, unlike other vendors, as a stand-alone product for investigators who prefer to use their own solutions and reagents.

 

Cell permeable, acetoxymethyl group (AM) esters

 

The native forms of K⁺-sensitive and Tl⁺-sensitive fluorescent indicators are negatively charged and membrane impermeable. However, masking the negative charge using non-polar, ester-linked moieties (AM esters) allow the molecules to enter cells through passive diffusion. Once inside the cell, ubiquitous intracellular esterase enzymes promote rapid hydrolysis of the AM esters leaving the active, highly polar form of the indicator trapped inside the cell.

 

Reagents for improving indicator loading and retention

 

Although the AM forms of ION’s fluorescent K⁺ and Tl⁺ indicators readily cross the plasma membrane, there are other factors that can affect how well the indicators load into cells. Because AM forms of the indicators are quite hydrophobic, it is common to use the non-ionic surfactant, Pluronic F-127, to improve their solubility in aqueous solution thus improving their loading into cells. Pluronic F-127 at the concentrations employed for loading indicators into cells is generally well tolerated. While Pluronic F-127 isn’t absolutely essential for loading most fluorescent K⁺ and Tl⁺ indicators into cells, it is highly recommended.

Another factor that can affect cell loading and intracellular localization is the presence of transport proteins capable of transporting many ion-sensitive, fluorescent indicators. In many cell types, the activity of these transporters can result in compartmentalization of the indicators into intracellular compartments as well as extrusion of the indicators outside of the cell. Both of these processes can dramatically reduce the signal-to-background levels and in some cell types (e.g. CHO cells),  make adequate loading of K⁺-selective fluorescent indicators nearly impossible. Fortunately, inhibitors of these transport proteins can be an effective means of improving cytoplasmic localization and indicator retention. ION offers a convenient, 100X ION-Pro inhibitor solution that may be used in conjunction with any of our K⁺-sensitive fluorescent indicators.

Be Brilliant! Explore our line of Potassium Indicators

Our collection of membrane permeable and impermeable ion indicators targeting K⁺.

 

Product Description Ex/Em (nm) Kd (nM) Solubility Filter Set
Brilliant Thallium Express High-throughput, no wash thallium (Tl⁺) assay. An optimal solution for measuring flux through potassium (K⁺), sodium (Na⁺), and non-selective cation channels and their effectors including transporters and GPCRs. 100 plates 490/515 FITC, GFP
Brilliant Thallium Flex High-throughput, no wash thallium (Tl⁺) assay. An optimal solution for measuring flux through potassium (K⁺), sodium (Na⁺), and non-selective cation channels and their effectors including transporters and GPCRs. 10 plates 490/515 FITC, GFP
IPG-1 AM Yellow-green fluorescent, potassium (K⁺) indicator, membrane permeable. Lower K⁺ affinity than IPG-2 and IPG-4. 525/545 50 DMSO FITC, GFP, YFP
IPG-1 TMA⁺ Salt Yellow-green fluorescent, potassium (K⁺) indicator, membrane impermeable. Lower K⁺ affinity than IPG-2 and IPG-4. 525/545 50 H20 FITC, GFP, YFP
IPG-2 AM Yellow-green fluorescent, potassium (K⁺) indicator, membrane permeable. Lower K⁺ affinity than IPG-4 and higher affinity than IPG-1 525/545 18 DMSO FITC, GFP, YFP
IPG-2 TMA+ SALT Yellow-green fluorescent, potassium (K⁺) indicator, membrane impermeable. Lower K+ affinity than IPG-4 and higher affinity than IPG-1 525/545 18 H20 FITC, GFP, YFP
IPG-4 AM Yellow-green fluorescent, potassium (K⁺) indicator, membrane permeable. Higher K⁺ affinity than IPG-1 and IPG-2 525/545 7 DMSO FITC, GFP, YFP
IPG-4 TMA+ SALT Yellow-green fluorescent, potassium (K⁺) indicator, membrane impermeable. Higher K⁺ affinity than IPG-1 and IPG-2 525/545 7 H20 FITC, GFP, YFP
PBFI Ratiometric potassium (K⁺) indicator, membrane permeable 340,380/505 5 DMSO Fura-2
PBFI K+ SALT Ratiometric potassium (K⁺) indicator, membrane impermeable 340,380/505 5 H20 Fura-2
THALLOS AM Best in class, green fluorescent, thallium (Tl⁺) indicator, membrane permeable. Works with potassium (K⁺), sodium (Na⁺), and monovalent cation channels, and transporters. 490/515 DMSO FITC, GFP

Sodium

Sodium indicators are molecules that exhibit an increase in fluorescence upon binding Na⁺.

Sodium (Na⁺) in physiology

Sodium, along with potassium (K⁺), is one of the most important monovalent metal cations in living organisms. Sodium channels, Na⁺-permeable non-selective monovalent cation channels, and Na⁺ transporters play critical roles including modulating neuronal activity, powering transport of nutrients and signaling molecules, and regulating solute balance. Na⁺-permeable channel and Na⁺ transporter-targeted drugs provide effective treatments for a diversity of indications: epilepsy, pain, bipolar disorder, depression, diuresis, and many others. An increasingly long list of Na⁺-permeable channels, and Na⁺ transporters are being associated with a wide variety of disorders. As a result, interest in Na⁺-permeable channels and Na⁺ transporters as drug targets remains high.

Because expanding our understanding of Na⁺ permeable channels and Na⁺ transporters are of such intense interest to both basic and drug discovery researchers, a facile means to measure their activity is required. Unfortunately, measuring intracellular and extracellular Na⁺ levels and dynamics using fluorescent indicators is challenging. In most cells at rest, cytoplasmic Na⁺ concentrations are ~5 mM while K⁺ concentrations are above 100 mM. Conversely, extracellular Na⁺ concentrations are typically >100 mM while K⁺ concentrations are in the 3-5 mM range. Therefore, an effective Na⁺ sensor needs to be able to discriminate between K⁺, Na⁺ and other common physiological cations while possessing an appropriate Na⁺ affinity to detect physiologically relevant changes in Na⁺ concentrations.

ING-2

ION Natrium Green-2 (ING-2), formerly known as Asante Natrium Green (ANG-2), is a small, synthetic fluorochrome fused with a Na⁺-binding moiety. Under conditions where Na⁺ is not bound, the fluorescence of the sensor is significantly quenched. When Na⁺ is bound, the quenching is relieved, and the fluorescence of the sensor dramatically increases.

ING-2 (EX 525 nm, EM 545 nm, Kd 20 mM) is the best-in-class fluorescent Na⁺ sensor replacing previous Na⁺ indicators such as SBFI and CoroNa Green. Its affinity for Na⁺ is well suited to respond to changes in intracellular Na⁺ concentrations resulting from activation of plasma membrane Na⁺ permeable channels and Na⁺ transporters. It is compatible with a wide variety of detectors including fluorescent microscopes, plate readers, and flow cytometers using common filter sets like sets (e.g. YFP and FITC) as well as multiphoton approaches. ING-2 has also proven useful for high-throughput screening (HTS).

Brilliant Sodium Assays

Brilliant Sodium Assays are the first kits designed to measure intracellular sodium in an HTS-compatible format, so you can discover and characterize the effects of many tens of thousands of compounds and environmental factors on effectors of intracellular Na⁺

While ING-2 is an excellent Na⁺-sensitive, fluorescent indicator, the challenges of discriminating between K⁺ and Na⁺ are not trivial, and together with the often modest changes in intracellular Na⁺ results in relatively small signals compared to those observed with the Ca2⁺-sensitive fluorescent indicators, such as Fluo-4.

In many instances, the Tl⁺ flux assay offers an outstanding alternative to ING-2 and other Na⁺-sensitive fluorescent indicators. The Tl⁺ flux assay takes advantage of a Tl⁺-selective fluorescent indicator, Thallos, and the fact that Na⁺ permeable channels and Na⁺ transporters readily accept Tl⁺ as a Na⁺ surrogate. When Tl⁺ is added to the outside of cells loaded with Thallos, Tl⁺ entering cells through Na⁺ permeable channels and Na+ transporters results in a dramatic increase in fluorescence. Although the Tl⁺ assay is most commonly associated with high-throughput screening (HTS) of K⁺ channels, it is equally suitable for HTS of Na⁺ permeable channels and Na⁺ transporters.

Thallos

(EX 490 nm, EM 515 nm) is the best-in-class Tl⁺-sensitive fluorescent indicator. It offers outstanding cell loading and excellent compatibility with commonly available FITC filters.

We provide ING-2 in membrane permeable and membrane impermeable forms, as well as our Brilliant Sodium Assays. We offer Thallos as part of our convenient Brilliant Thallium Assay kits and, unlike other vendors, as a stand-alone product for investigators who prefer to use their own solutions and reagents.

 

Cell permeable, acetoxymethyl group (AM) esters

 

The native forms of the Na⁺-sensitive and Tl⁺-sensitive fluorescent indicators are negatively charged and membrane impermeable. However, masking the negative charge using non-polar, ester-linked moieties (AM esters) allow the molecules to enter cells through passive diffusion. Once inside the cell, ubiquitous intracellular esterase enzymes promote rapid hydrolysis of the AM esters leaving the active, highly polar form of the indicator trapped inside the cell.

 

Reagents for improving indicator loading, retention and background suppression

 

Although the AM forms of ION’s fluorescent, Na⁺ and Tl⁺ indicators readily cross the plasma membrane, there are other factors that can affect how well the indicators load into cells. Because AM forms of the indicators are quite hydrophobic, it is common to use the non-ionic surfactant, Pluronic F-127, to improve their solubility in aqueous solution thus improving their loading into cells. Pluronic F-127 at the concentrations employed for loading indicators into cells is generally well tolerated. While Pluronic F-127 isn’t absolutely essential for loading most fluorescent Na⁺ and Tl⁺ indicators into cells, it is highly recommended.

Another factor that can affect cell loading and intracellular localization is the presence of anion transport proteins in both plasma and intracellular membranes. In many cell types, the activity of these transporters can result in compartmentalization of the indicators into intracellular compartments as well as extrusion of the indicators outside of the cell. Both of these processes can dramatically reduce the signal-to-background levels and in some cell types (e.g. CHO cells), make adequate loading of Na⁺ and Tl⁺ sensitive fluorescent indicators nearly impossible. Fortunately, probenecid, an organic anion transport inhibitor can be an effective means of inhibiting the transporters, improving cytoplasmic localization and indicator retention. ION offers a convenient, 100X probenecid solution that may be used in conjunction with our Na⁺ and Tl⁺-sensitive fluorescent indicators. While probenecid is generally well-tolerated over the time course of typical experiments using Na⁺ and Tl⁺-sensitive fluorescent indicators, effects on the activity of TRP channels has been reported.

Be Brilliant! Explore our line of Sodium Indicators

Our collection of membrane permeable and impermeable ion indicators targeting Na⁺.

 

Product Description Ex/Em (nm) Kd (nM) Solubility Filter Set
Brilliant Thallium Express High-throughput, no wash thallium (Tl⁺) assay. An optimal solution for measuring flux through potassium (K⁺), sodium (Na⁺), and non-selective cation channels and their effectors including transporters and GPCRs. 100 plates 490/515 FITC, GFP
Brilliant Thallium Flex High-throughput, no wash thallium (Tl⁺) assay. An optimal solution for measuring flux through potassium (K⁺), sodium (Na⁺), and non-selective cation channels and their effectors including transporters and GPCRs. 10 plates 490/515 FITC, GFP
THALLOS AM Best in class, green fluorescent, thallium (Tl⁺) indicator, membrane permeable. Works with potassium (K⁺), sodium (Na⁺), and monovalent cation channels, and transporters. 490/515 DMSO FITC, GFP
Brilliant Sodium Assays High-throughput, no wash sodium (Na⁺) assay for measuring changes in intracellular Na⁺ caused by effectors of Na⁺ channels and transporters 525/545 FITC, GFP, YFP
IPG-2 AM Yellow-green fluorescent, sodium (Na⁺) indicator, membrane permeable. Higher Na⁺ affinity than ING-1 525/545 20 DMSO FITC, GFP, YFP
IPG-2 TMA+ SALT Yellow-green fluorescent, sodium (Na⁺) indicator, membrane impermeable. Higher Na⁺ affinity than ING-1 525/545 20 H20 FITC, GFP, YFP
IPG-4 AM Ratiometric sodium (Na⁺) indicator, membrane permeable 340, 380/505 4 DMSO Fura-2

Thallium

ION Biosciences Thallium Indicators

Labeled thallium indicators are molecules that exhibit an increase in fluorescence upon binding Tl⁺

HeLa cells loaded with Thallos in the presence (+) or the absence (-) of 0.5 mM Tl⁺

Thallium (Tl⁺)-sensitive fluorescent indicators

Sodium (Na⁺) and potassium (K⁺) are the two most physiologically important monovalent metal cations. Transporters and ion channels for Na⁺ and K⁺ are found in every cell of every known organism and are critical for life. While drugs targeting these channels and transporters are used to treat indications ranging from type II diabetes, to epilepsies, to blood pressure, the therapeutic potential of this incredibly diverse set of proteins remains largely unknown. Therefore, tools to promote the rapid discovery and evaluation of monovalent cation channels and transporters as potential therapeutic targets are badly needed.

While ION’s class-leading Na⁺ and K⁺-sensitive fluorescent indicators are excellent tools for studying cellular Na⁺ and K⁺ dynamics, the relatively small changes in the intracellular concentrations of Na⁺ and K⁺ under normal physiological conditions can make Na⁺ and K⁺ indicators challenging to use, particularly for high-throughput screening (HTS) applications. In many instances, the Tl⁺ flux assay offers an outstanding alternative to K⁺-sensitive and Na+-sensitive fluorescent indicators.

The Tl⁺ flux assay takes advantage of a Tl⁺-selective fluorescent indicator, Thallos, and the fact that K⁺ and Na⁺ permeable channels and K⁺ and Na⁺ transporters readily accept Tl⁺ as surrogate cation. When Tl⁺ is added to the outside of cells loaded with Thallos, Tl⁺ entering cells through ion channels and transporters permeable to Na⁺ or K⁺ results in a dramatic increase in fluorescence. Although Tl+ flux assay is the most commonly associated with HTS of K⁺ channels, it is equally suitable for HTS of Na⁺ permeable channels and Na+ transporters as well as non-selective, monovalent cation channels.

Thallos is a small, synthetic fluorochrome which incorporates a Tl⁺-binding moiety with excellent selectivity for Tl⁺ over most other physiologically relevant metal cations (e.g. Ca²⁺). Under conditions where Tl⁺ is not bound, the fluorescence of the sensor is significantly quenched. When Tl⁺ is bound, the quenching is relieved, and the fluorescence of the sensor dramatically increases.

Thallos is compatible with a wide variety of detectors including fluorescent microscopes, plate readers, flow cytometers. Over the last 20 years, since Dr. David Weaver invented the Tl⁺ flux assay, Tl⁺-sensitive indicators have found numerous uses including high-throughput screening (HTS) of K⁺ channel, Na⁺ channel, other monovalent cation channels, some monovalent cation transporters, and the Gi/o class of G protein-coupled receptors (GPCR) discovery.

Thallos

(EX 490 nm, EM 515 nm) is the best-in-class Tl⁺-sensitive fluorescent indicator. It offers outstanding cell loading and excellent compatibility with commonly available FITC filters.

Brilliant Thallium Flex

(EX 490 nm, EM 515 nm) High-throughput, no wash thallium (Tl⁺) assay. The most flexible solution for measuring the activity of potassium (K⁺), sodium (Na⁺), and non-selective cation channels and their effectors including transporters and GPCRs. 10 plates

Brilliant Thallium Express

(EX 490 nm, EM 515 nm) High-throughput, no wash thallium (Tl⁺) assay. A streamlined solution for large-scale screening of potassium (K⁺), sodium (Na⁺), and non-selective cation channels and their effectors including transporters and GPCRs. 100 plates

Cell permeable, acetoxymethyl group (AM) esters

The native form of Thallos is negatively charged and membrane impermeable. However, masking the negative charge using non-polar, ester-linked moieties (AM esters) allow the molecules to enter cells through passive diffusion. Once inside the cell, ubiquitous intracellular esterase enzymes promote rapid hydrolysis of the AM esters leaving the active, highly polar form of the indicator trapped inside the cell.

Reagents for improving indicator loading, retention and background suppression

 

Although the AM form of Thallos indicator readily crosses the plasma membrane, there are other factors that can affect how well Thallos loads into cells. Because Thallos-AM is quite hydrophobic, it is common to use the non-ionic surfactant, Pluronic F-127, to improve its solubility in aqueous solution thus improving its loading into cells. Pluronic F-127 at the concentrations employed for loading indicators into cells is generally well tolerated. While Pluronic F-127 isn’t absolutely essential for loading Thallos into cells, it is highly recommended.

Another factor that can affect cell loading and intracellular localization is the presence of transport proteins capable of transporting many ion-sensitive, fluorescent indicators. In many cell types, the activity of these transporters can result in compartmentalization of the indicators into intracellular compartments as well as extrusion of the indicators outside of the cell. Both of these processes can dramatically reduce the signal-to-background levels and in some cell types (e.g. CHO cells),  make adequate loading of Tl⁺-selective fluorescent indicators nearly impossible. Fortunately, inhibitors of these transport proteins can be an effective means of improving cytoplasmic localization and indicator retention. ION offers a convenient, 100X ION-Pro inhibitor solution that may be used in conjunction with any of our Tl⁺-sensitive fluorescent indicators.

Be Brilliant! Explore our line of Thallium Indicators

Our collection of ion indicators targeting Tl⁺ flux.

 

Product Description Ex/Em (nm) Kd (nM) Solubility Filter Set
Brilliant Thallium Express High-throughput, no wash thallium (Tl⁺) assay. An optimal solution for measuring flux through potassium (K⁺), sodium (Na⁺), and non-selective cation channels and their effectors including transporters and GPCRs. 100 plates 490/515 FITC, GFP
Brilliant Thallium Flex High-throughput, no wash thallium (Tl⁺) assay. An optimal solution for measuring flux through potassium (K⁺), sodium (Na⁺), and non-selective cation channels and their effectors including transporters and GPCRs. 10 plates 490/515 FITC, GFP
THALLOS AM Best in class, green fluorescent, thallium (Tl⁺) indicator, membrane permeable. Works with potassium (K⁺), sodium (Na⁺), and monovalent cation channels, and transporters. 490/515 DMSO FITC, GFP

Cell Health

Our best tools for cellular health to advance your research

An essential step in early stage, translational research is measuring cytotoxicity. ION’s product portfolio is designed to help you easily capture important parameters of cell health, whether your preferred instrument is a plate reader, fluorescence microscope, or flow cytometer, using a collection of criteria that are indicative metrics of cell viability or death. These criteria include intracellular esterase activity as a marker of cell viability, plasma membrane integrity as a marker of cell death, and mitochondrial membrane potential as an early-stage marker of apoptosis. When used in combination, these assays can provide an informative snapshot of cell health.

Intracellular enzyme activity and dye retention is a hallmark of viable cells. During cell death, increased plasma membrane permeability causes enzyme leakage and a reduction in activity. Dyes that measure esterase activity are modified with ester-moieties, which help them passively diffuse through cell membranes and render the dyes non-fluorescent. Once inside the cell, ubiquitous esterases present in the cytosol cleave ester-moieties, activating dye fluorescence and trapping the dye within the cell. ION sells Calcein AM in multiple formats (dry or in a prepared solution) and in combination with Ethidium Homodimer I as part of ION Vital – Viability so you can simultaneously detect dead cells.

Calcein AM

(Ex/Em 495 nm/515 nm) – Calcein AM is a membrane-permeant, non-fluorescent form of calcein that is converted to green fluorescent calcein in viable cells, resulting in uniform cytosolic fluorescence. Calcein is well retained within the cytosol of most healthy cells with intact cell membranes.

BCECF AM

(Ex/Em 490 nm/535 nm) – BCECF AM is the most popular green fluorescent, intracellular pH indicator, however it can also be used as a marker of intracellular esterase activity. For this application, BCECF should be used in non-ratiometric mode using standard FITC excitation and emission settings.

ION Vital – Viability Kit

This kit combines Ethidium Homodimer I and Calcein AM to provide a two-color, fluorescence-based assay to discriminate between live and dead cells.

A loss of plasma membrane integrity is an indicator of cell death. Nucleic acid-binding, cell impermeant dyes, like Ethidium Homodimer I (EthD-I), can be used to label the nuclei of cells with compromised plasma membranes. These dyes are excluded from intact, healthy cells and exhibit minimal fluorescence in the absence of nucleic acids. Once the dye crosses the cell membrane and binds to nucleic acids (DNA) inside the cell, it displays a significant increase in fluorescence. ION sells EthD-I in multiple formats (dry or in a prepared solution) and in combination with Calcein AM as part of ION Vital – Viability so you can simultaneously detect viable cells.

Ethidium Homodimer I

(Ex/Em: 528 nm/617 nm) – EthD-I is a membrane-impermeable, high-affinity, nucleic acid stain that selectively labels dead cells with compromised plasma membranes.

ION Vital – Viability Kit

This kit combines Ethidium Homodimer I and Calcein AM to provide a two-color, fluorescence-based assay to discriminate between live and dead cells.

Mitochondrial membrane potential is one of the driving forces behind mitochondrial ATP synthesis, and is necessary for maintaining cell homeostasis. During early stages of apoptosis, mitochondrial function and membrane integrity are disrupted by the activation of apoptotic proteins and subsequent release of cytochrome C, mitochondrial Ca2+ stores, and other components. Membrane potential sensitive dyes, such as JC-1 and JC-10, can be used to monitor the loss of mitochondrial membrane polarization observed during apoptosis. These dyes compartmentalize in healthy mitochondria, leading to the formation of aggregates that exhibit orange fluorescence. When mitochondrial membrane potential is lost during apoptosis, these dyes no longer accumulate and return to green fluorescent monomers. ION sells JC-10 individually and as part of our optimized ION Vital Assay line of products

JC-10

(Ex/Em: 490,540 nm/525, 590nm) – JC-10 is a mitochondrial membrane potential probe. It possesses superior aqueous solubility compared to its better known analogue, JC-1. At low concentrations, JC-10 is monomeric and emits a green fluorescence. JC-10 accumulates in healthy mitochondria, forming J-aggregates that exhibit an orange fluorescence. Its potentiometric behavior allows for ratiometric analysis of mitochondrial membrane potential, where a shift from orange to green fluorescence is indicative of compromised mitochondria.

ION Vital – MitoVolt

MitoVolt is a mitochondrial membrane potential assay kit that uses JC-10, a potentiometric dye that is more soluble than JC-1. It is an ideal solution for detecting changes in mitochondrial membrane potential due to cell apoptosis or other stress-inducing phenomena, and is compatible with fluorescence microscopy, flow cytometry, and plate reader applications.

Be Brilliant! Explore our line of products for Cell Health

 

Product Description
Calcein AM Green fluorescent, live cell indicator.
Ethidium Homodimer I Red fluorescent, dead cell stain
ION VITAL – Viability Kit Two-color, fluorescence-based assay to discriminate between live and dead cells
ION VITAL – MitoVolt Mitochondrial membrane potential assay kit that uses JC-10, a membrane potential sensitive dye that is more soluble than JC-1.

G protein-coupled receptor (GPCR)

G protein-coupled receptor (GPCR) assay technologies

GPCRs (aka 7-transmembrane receptors) are a large and diverse class of transmembrane signaling proteins. They have tremendously diverse functions ranging from neurotransmitter receptors to cellular adhesion. GPCRs mediate their physiological effects in a variety of ways, most famously by coupling to downstream effectors through the heterotrimeric G protein complex (Gα,β,γ) but also through other pathways such as β-arrestins. GPCRs have been very effectively targeted for therapeutics – making up ~30% of all approved drugs. Over the last decade-and-a-half, allosteric modulation of GPCRs and unanticipated pharmacological diversity has led to a new generation of intense interest in GPCRs as therapeutic targets, therefore rapid and effective ways of measuring GPCR activation are critically important.

A key signaling pathway for many GPCRs is via Gaq-mediated activation of phospholipase C-β resulting in generation of inositol 1,4,5-trisphosphate (IP3) and subsequent activation of IP3 receptor Ca²⁺ channels causing release of Ca²⁺ from intracellular stores. Using fluorescent, Ca²⁺-sensitive indicators, this signaling pathway has been exploited for decades as a rapid and effective means of measuring GPCR activation

 

ION offers a variety of fluorescent, Ca²⁺-sensitive indicators that are ideal for measuring GPCR activation using microscopy, flow cytometry, or plate readers.

Fluo-4 

(EX 490 nm/Em 515 nm, Kd 355 nM) is the most used and the most trusted of the single-wavelength Ca²⁺ indicators. It has been the indicator of choice for countless basic research studies and has been used to screen many tens-of-millions of compounds using high-throughput screening (HTS) to discover pharmacological tools and drugs for GPCRs, and a wide variety of Ca²⁺ permeable ion channels and transporters.

Fluo-2 

(EX 490 nm/Em 515 nm, Kd 290 nM) Also known as Fluo-2 HA, Fluo-2 offers the same spectral properties as Fluo-4 combined with superior loading in many cell types as well as a slightly higher Ca²⁺ affinity. The improved loading and increased affinity can result in superior sensitivity.

Brilliant Calcium Express Kit

Our Express kit is custom-tailored to customer’s needs, providing only the reagents you need in the amounts you need them. Less waste, higher reproducibility, and more convenience.

Brilliant Calcium Express is a (EX 490 nm, EM 515 nm) High-throughput, no wash calcium (Ca²⁺) assay. The most flexible solution for measuring the activity of GPCRs coupled to intracellular Ca²⁺ release. 10 plates

Brilliant Calcium Flex Kit

Our Express kit is custom-tailored to customer’s needs, providing only the reagents you need in the amounts you need them. Less waste, higher reproducibility, and more convenience.

Brilliant Calcium Express is a (EX 490 nm, EM 515 nm) High-throughput, no wash calcium (Ca²⁺) assay. The most flexible solution for measuring the activity of GPCRs coupled to intracellular Ca²⁺ release. 10 plates

A key signaling pathway for another major class of GPCRs, Gi/o-coupled GPCRs, is through activation of G protein-gated Inward Rectifying Potassium (K⁺) channels, better known as GIRK channels. Not only is the GPCR-mediated modulation of GIRK channels of great physiological importance, it also provides a convenient means of measuring Gi/o GPCR activity using Tl⁺ flux. This GIRK assay approach offers some key advantages compared to other approaches used to measure the activity of Gi/o GPCRs. Unlike the most common approaches which utilize fixed-endpoint measures of cAMP, the GIRK assay is a real-time measure of GPCR activation and does not require non-physiological elevations of intracellular cAMP. Unlike approaches which utilize chimeric Gα subunits (e.g. Gqi5) to induced Gi/0 GPCRs to promote intracellular Ca2+ release, the GIRK assay enables measurement of Gi/o GPCR activity through their native Gβ/γ coupling mechanism.

ION offers Tl⁺ flux reagents and assay kits to help enable researchers to utilize GIRK-based GPCR activity assays.

Brilliant Thallium Express

(EX 490 nm, EM 515 nm) High-throughput, no wash thallium (Tl⁺) assay. A streamlined solution for large-scale screening of potassium (K⁺), sodium (Na⁺), and non-selective cation channels and their effectors including transporters and GPCRs. 100 plates

Brilliant Thallium Flex

(EX 490 nm, EM 515 nm) High-throughput, no wash thallium (Tl⁺) assay. The most flexible solution for measuring the activity of potassium (K⁺), sodium (Na⁺), and non-selective cation channels and their effectors including transporters and GPCRs. 10 plates

Thallos

(EX 490 nm, EM 515 nm) is the best-in-class Tl⁺-sensitive fluorescent indicator. It offers outstanding cell loading and excellent compatibility with commonly available FITC filters.

Be Brilliant! Explore our line of GPCRs Assays

Our collection of G protein-coupled receptor assay technologies

 

Product Description Ex/Em (nm) Kd (nM) Solubility Filter Set
Brilliant Thallium Express High-throughput, no wash thallium (Tl⁺) assay. An optimal solution for measuring flux through potassium (K⁺), sodium (Na⁺), and non-selective cation channels and their effectors including transporters and GPCRs. 100 plates 490/515 FITC, GFP
Brilliant Thallium Flex High-throughput, no wash thallium (Tl⁺) assay. An optimal solution for measuring flux through potassium (K⁺), sodium (Na⁺), and non-selective cation channels and their effectors including transporters and GPCRs. 10 plates 490/515 FITC, GFP
THALLOS AM Best in class, green fluorescent, thallium (Tl⁺) indicator, membrane permeable. Works with potassium (K⁺), sodium (Na⁺), and monovalent cation channels, and transporters. 490/515 DMSO FITC, GFP
Brilliant Calcium Flex High-throughput, no wash calcium (Ca²⁺) assay. An optimal solution for measuring intracellular Ca²⁺ dynamics, including effectors such as GPCRs and protein transporters. 10 plates FITC, GFP
Brilliant Calcium Express High-throughput, no wash calcium (Ca²⁺) assay. An optimal solution for measuring intracellular Ca²⁺ dynamics, including effectors such as GPCRs and protein transporters. 100 plates FITC, GFP
ION Essentials Ratiometric Calcium The necessary reagents for conducting no wash, ratiometric calcium flux assays compatible with plate reader and fluorescence microscopy applications. Individual components are provided to give the user the flexibility needed to customize your assay. DMSO GFP/FITC. PI/Texas Red
Fluo-4 AM Green fluorescent, calcium (Ca²⁺) indicator, membrane permeable 145 DMSO FITC. GFP
Fluo-3 AM Green fluorescent, calcium (Ca²⁺) indicator, membrane permeable Green fluorescent, calcium (Ca²⁺) indicator, membrane permeable 355 DMSO FITC
Fluo-2 AM Green fluorescent, calcium (Ca²⁺) indicator, membrane permeable. Higher Ca²⁺ affinity than Fluo-4. 390 DMSO FITC, GFP
Fura-2 AM UV-excitable, ratiometric green calcium (Ca²⁺) indicator, membrane permeable 290 DMSO Fura-2
ICR-1 AM Red fluorescent, calcium (Ca²⁺) indicator, membrane permeable. Optimal for multiplexing and tissue imaging. Fluorescence lifetime and multiphoton compatible. Green fluorescent, calcium (Ca²⁺) indicator, membrane permeable. Higher Ca²⁺ affinity than Fluo-4. 480 DMSO Texas Red

High Throughput Screening

ION Biosciences’ Brilliant Calcium Assay series is a total assay solution for multi-well plate-based, high-throughput measurements of dynamic changes in intracellular Ca²⁺. The Flex and  Express kits represent different sizes and packaged configurations for various screening scenarios.  Each assay kit is designed to be a simple, fast, and reliable fluorescence-based assay.

Whether your platform is FLIPR®,  FlexStation®, or FDSS, ION’s assay kits will provide a  smooth process for your assay development.

There are no intermediate wash-steps required with these kits. In each version, you will find all the reagents necessary for preparation to use as a no-wash or washed assays for adherent and non- adherent  cells.

Brilliant Sodium Assays

High-throughput, no wash sodium (Na⁺) assay for measuring changes in intracellular Na⁺ caused by effectors of Na⁺ channels and transporters

SPECIFICATIONS

Excitation Emission MW:
525nm 545nm 1084

 

Product Code: Product Description
9000-10 Brilliant Sodium Flex
9000-100 Brilliant Sodium Express

PRODUCT DESCRIPTION

ION’s Brilliant Sodium Assay is a total assay solution for multi-well plate-based, high-throughput measurements of changes in intracellular Na⁺ mediated through a wide-variety of plasma membrane and intracellular sodium channels and transporters. In multi-well, plate-based formats, the Brilliant Sodium Assay can be used to discover and characterize the effects of many tens-of-thousands of compounds and environmental factors on effectors of intracellular Na⁺. ION’s Brilliant Sodium Assay provides all the reagents necessary for use as a wash or no-wash assay with adherent or non-adherent cells. The optional use of a probenecid solution and an extracellular background masking solution (TRS) offers the ultimate in compatibility for cells types which are difficult to load with fluorescent Na⁺ indicators (e.g. Chinese Hamster Ovary, CHO cells) and when performing assays in complete, serum-containing cell culture medium is desired.

ION’s Brilliant Sodium Assay is compatible with fluorescence microscopes, flow cytometers, and plate readers capable of detecting fluorescein or more optimally, yellow fluorescent protein (YFP).

Our Flex kit provides all necessary reagents in a format to maximize flexibility for your assay design and customization challenges. If you want a kit designed just for you, inquire about our Brilliant Sodium Express Kit.

REFERENCES

Laila Blömer, Marco Canepari, Luiza Filipis. Ultrafast Sodium Imaging of the Axon Initial Segment of Neurons in Mouse Brain Slices. Current Protocols in Bioinformatics, Wiley, 2021, 1 (3). https://doi.org/10.1002/cpz1.64

Tay B, Stewart TA, Davis FM, Deuis JR, Vetter I. Development of a high-throughput fluorescent no-wash sodium influx assay. PLoS One. 2019 Mar 11;14(3):e0213751.

Iamshanova, O., Mariot, P., Lehen’kyi, V. et al. Comparison of fluorescence probes for intracellular sodium imaging in prostate cancer cell lines. Eur Biophys J. 45, 765–777 (2016).

Yurinskaya VE, Aksenov ND, Moshkov AV, Goryachaya TS, Vereninov AA. Fluorometric Na+ Evaluation in Single Cells Using Flow Cytometry: Comparison with Flame Emission Assay. Cell Physiol Biochem. 2020 May 29;54(4):556-566.

ION Biosciences Brilliant Thallium Assay is a total assay solution for multi-well plate-based, high-throughput measurements of Tl⁺ flux through K⁺, Na⁺, non-selective cation channels, and some Na⁺ or K⁺ transporters. The ION Biosciences Brilliant Thallium Assay is also useful for a wide range of effectors of ion channels and transporters including G protein-coupled receptors, lipid and protein kinase, for example. In multi-well, plate-based formats, the Brilliant Thallium Assay can be used to discover and characterize the effects of tens to many tens-of-thousands of compounds and environmental factors on effectors of Tl⁺ flux.

ION VITAL – Viability Kit

Two-color, fluorescence-based assay to discriminate between live and dead cells

SPECIFICATIONS

Excitation Emission
Live 495 nm   Dead 528 nm  Live 515 nm   Dead 617 nm 

 

Product Code: Product Description
5000 ION VITAL – Viability Kit

PRODUCT DESCRIPTION

ION Vital – Viability kit can be used to quantify live and dead cells within eukaryotic cell suspensions or adherent cultures, 3D cultures, organoids, and some non-fixed tissues. Cannot be used for yeast or bacteria. When following our protocol, ION Vital—Viability kit provides enough reagents to make 150 mL of working solution, enough for fifteen 96-well plates or 1,500 flow cytometry samples. The actual number of assays will vary according to optimal dye concentrations for your application.

ION VITAL – MitoVolt

Mitochondrial membrane potential assay kit that uses JC-10, a membrane potential sensitive dye that is more soluble than JC-1.

SPECIFICATIONS

 

Excitation Emission MW: Solubility: Purity (minimum):
490 540nm 525 590nm ~600 DMSO or H2O >98%

 

Product Code: Product Description
5100 ION VITAL – MitoVolt

PRODUCT DESCRIPTION

ION Vital – MitoVolt assay kit is the ideal solution for detecting changes in mitochondrial membrane potential due to cell apoptosis or other stress-inducing phenomena. ION Vital – MitoVolt is compatible with fluorescence microscopy, flow cytometry, and plate reader applications.

JC-10 is a mitochondrial membrane potential probe. It possesses superior aqueous solubility compared to its better known analogue, JC-1. At low concentrations, JC-10 is monomeric and emits a green fluorescence. JC-10 accumulates in healthy mitochondria, forming J-aggregates that exhibit an orange fluorescence. Mitochondrial depolarization, a key marker of cellular apoptosis, results in a loss of JC-10 accumulation and a reversal to its monomeric form. This reversible behavior of JC-10 allows for ratiometric analysis of mitochondrial membrane potential, where a shift from orange (Ex/Em: 540nm/590nm) to green fluorescence (Ex/Em: 490nm/525nm) is indicative of compromised mitochondria.

ION Vital – MitoVolt can be used to monitor and/or visualize mitochondrial membrane potential as a static endpoint or in real-time. When following our protocol, ION Vital – MitoVolt provides enough reagents to make 25 mL of working solution, enough for five 96-well plates or 500 flow cytometry samples. The actual number of assays will vary according to optimal dye concentrations and assay volumes for your application.

Multidrug Resistance Activity Kit

Two-color, fluorescence-based assay to discriminate between live and dead cells

SPECIFICATIONS

Excitation Emission
495nm 515nm

 

Product Code: Product Description
3330 Multidrug Resistance Activity Kit

PRODUCT DESCRIPTION

ION’s Multidrug Resistance (MDR) Activity kit is an effective solution for detecting MDR1 and MRP1 activity and compounds susceptible to MDR-mediated efflux. ION’s MDR Activity kit is compatible with fluorescence microscopy, flow cytometry, and fluorescence plate readers using FITC/fluorescein settings.

Calcein AM is a membrane-permeant, non-fluorescent dye that enters cells passively. Once inside the cytosol of cells, intracellular esterases convert it to fluorescent Calcein (Ex/Em: 495 nm/515 nm), resulting in uniform cytosolic fluorescence. Drug efflux transporters, such as P-glycoprotein (Pgp, MDR1) and multidrug-resistance-associated protein (MRP1), actively extrude Calcein AM from inside the cell before esterases can convert it to Calcein. The presence of additional MDR substrates or inhibitors of MDR expression results in decreased Calcein AM efflux, causing a measurable increase in intracellular fluorescence. In addition to identifying MDR substrates and inhibitors, this kit can also be used to evaluate the activity of MDR transporters in cells.
When following our protocol, ION’s MDR Activity kit provides enough reagents to make 100 mL of working solution, enough for ten 96- or 384-well plates or 80 flow cytometry samples. The actual number of assays will vary according to optimal dye concentrations for your application.

Custom Services

Assay Development

ION scientists have over 2 decades of experience using fluorescent, metal ion-sensitive indicators to develop and validate high-throughput screening compatible (HTS) assays for a broad diversity of ion channels, ion transporters and G protein-coupled receptors.

Free up valuable resources by outsourcing your development needs to ION.

We would love to be your source for the highest quality custom assays.

Please, contact us to learn how we can help further your research goals: info@stratech.co.uk