Calcium Visualization For All Platforms

Calcium is a ubiquitous cellular messenger impacting a number of signal transduction pathways, with multiple roles in processes ranging from neurotransmission to muscle contraction. Single-channel indicators, ratiometric probes, and calcium flux assays to track calcium mobilization are all in perpetually high demand. AAT Bioquest are proud to have developed the largest collection of calcium indicators, including well-known favorites Fluo-8® AM, Cal-520® AM, and Calbryte™ 520 AM. We have also expanded our offerings of calcium flux assays and indicator conjugates, with intense fluorescent signals and in a variety of wavelength options ranging from ultraviolet to infrared.

Fluo 8® Calcium Indicators

Fluo-8® AM addresses one of the major limitations common to fluo-based dyes, namely, the harsh conditions required for cell loading. Traditional fluo-based dyes such as Fluo- 2 AM, Fluo-3 AM and Fluo-4 AM need to be loaded and incubated with cells at 37°C for up to an hour before a reading can be taken. This not only subjects cells to harsh temperatures, but also requires users to spend time heating and cooling cell cultures. Additionally, it introduces incubation temperature as a possible confounding variable. Fluo-8® AM resolves all these problem by providing a robust calcium indicator that can be loaded into cells at room temperature. Fluo-8® AM is less temperature dependent than other fluo-based dyes, allowing for more consistent results under a wider range of experimental conditions. Furthermore, with an incubation time of as little as 20 minutes, Fluo-8® AM enables fast turnaround for improved productivity. The best part is that even with structural modifications and improvements, Fluo-8® AM’s spectral properties remains unchanged, maintaining the familiar spectra of Fluo-4 AM (ie. Ex/Em = 494/517nm).

Key features of Fluo 8® calcium indicators

  • Faster cell loading: Fluo 8® can be readily loaded into cells at room temperature. Fluo-3 AM and Fluo-4 AM require a loading temperature of 37°C
  • Brighter fluorescence: Fluo 8® is 2 times brighter than Fluo-4 AM, and 4 times brighter than Fluo-3 AM
  • Convenient: Fluo 8®’s identical spectra to those of Fluo-4 and Fluo-3 makes upgrading easy

Cal-520® Calcium Indicators

The Cal-520® calcium indicators series is the most comprehensive portfolio of fluorogenic calcium-sensitive dyes. It includes six unique indicators, Cal-500™, Cal-520®, Cal-590™, Cal-630™, Cal-670™ and Cal-770™, which differ primarily in their absorbance and emission wavelengths. Each of the indicators emit minimal fluorescence at resting levels of calcium, and increases its fluorescence intensity >100-fold at saturating calcium concentrations. The Cal-520&reg series of indicators are available as membrane-permeant AM esters, and membrane-impermeant salt derivatives and dextran conjugates.

Key features of Cal-520® calcium indicators

  • Higher signal-to-background ratios than Fluo-3, Fluo-4, Rhod-2 and Rhod-5
  • Enhanced intracellular retention
  • Convenient and efficient cell loading
  • Homogenous cytosolic localization
  • Red-shifted Cal-590™, Cal-630™, Cal-670™ and Cal-770™ can be used with GFP or other green-fluorescent dyes for multiplexing
  • Cal-770™ may be used for in vivo studies
  • Available as membrane-permeant AM esters (compatible with imaging, flow cytometry and microplate assays, such as HTS)
  • Available as membrane-impermeant salt derivatives for loading by microinjection, patch pipette, or pinocytic loading
  • Available as reactive-dyes for conjugation to a carrier

Calbryte™ Calcium Indicators

The Calbryte™ series is a family of the brightest and most robust calcium indicators developed to monitor intracellular calcium mobilization. It includes three novel calcium indicators, Calbryte™ 520, Calbryte™ 590 and Calbryte™ 630, which differ primarily in their absorbance and emission wavelengths. Calbryte™ probes emit fluorescence signals at significantly higher signal-to-background characteristics, and have greatly enhanced cellular retention. The Calbryte™ series has been optimized for use with fluorescence microscopes, fluorescence microplate readers and flow cytometers. They can also be used in high-throughput screening applications.

Key features of Calbryte™ calcium indicators

  • Calbryte™ probes are significantly brighter than any other calcium indicators under the same conditions
  • Greatly improved signal-to-background ratios
  • Enhanced intracellular retention, eliminates the use of probenecid
  • Fast and convenient cell loading in 45 to 60 minutes
  • Homogenous cytosolic localization
  • Red-shifted Calbryte™ 590 and Calbryte™ 630 can be used with GFP or other green-fluorescent dyes for multiplexing
  • Available as membrane-permeant AM esters (compatible with imaging, flow cytometry and microplate assays, such as HTS)
  • Available as membrane-impermeant salt derivatives for loading by microinjection, patch pipette, or pinocytic loading
  • Available as a reactive-dye for conjugation to a carrier