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Cal-520®, AM
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Cal-520®, AM：
Cal-520® AM is a next-generation fluorescent calcium indicator with an Ex/Em of 492/514 nm. It has several critical advantages and improvements over older generations of fluorescent Ca2+ probes such as Fluo-3 AM and Fluo-4 AM, namely:
The History
Every single year, there are hundreds of thousands of peer-reviewed papers published focusing on calcium and its role in biological systems. The reason for this is clear. Calcium ions (Ca2+) play a critical part in many biological processes, from the contracting of muscles to the release of neurotransmitters. This pervasiveness of calcium in biology, and the subsequent need to study it, has pushed for the development of viable, convenient calcium indicators, tools that researchers may use to quantify and visualize intracellular calcium.
The first big breakthrough came in the 1980s, when a team of scientists at the University of California Berkeley developed a fluorescein-based calcium indicator. Their research paved the way for now well-established calcium probes such as Fluo-3 AM and Fluo-4 AM. These fluorescein-based AM esters readily pass through the membranes of living cells, where once inside, they are rapidly hydrolyzed by esterases and set to detect calcium. Upon successful binding with calcium, Fluo-3 or Fluo-4 will become fluorescent, emitting a signal at ~520 nm when excited by a ~490 nm light source.
A significant achievement in its own right, these first generation fluorescein-based dyes did however suffer from several major drawbacks. One of these drawbacks was the high background fluorescence, sometimes termed noise. In general, noise creates problems in data analysis and can obscure possibly significant results, and should be minimized as much as possible. That is why we set out to synthesize a new, low noise fluorescent calcium indicator. After much research and optimizing by our dedicated team of experts, we have developed a new high signal to noise ratio calcium indicator, Cal-520® AM.
Cal-520® AM Features
Cal-520® AM is a next-generation fluorescent calcium indicator that has significant improvements over Fluo-3 AM and Fluo-4 AM. First, by improving the binding specificity of Cal-520® for calcium ions, we have managed to produce an indicator which reduces autofluorescence. This simultaneously lowers background fluorescence and raises the signal to noise ratio. The binding of Cal-520® to Ca2+ will result in a greater than one hundred fold increase in fluorescence intensity. Additionally, we have optimized Cal-520®'s chemical structure so that it is better localized. Unlike Rhod-2, which will localize in mitochondria, Cal-520® will predominantly localize in the cytosol. Lastly, we have developed Cal-520® AM for easy use in multiplexing. While Cal-520® AM retains the familiar spectra of Fluo-4 AM and Fluo-8® AM, we also have Cal-590™ AM (Ex/Em = 573/588 nm) and Cal-630™ (Ex/Em = 608/626 nm) for longer wavelength excitations.
Table 1. Various Excitations and Emissions for Multiplex Assays
Because of Cal-520® AM's excellent chemical properties, it is a robust probe for fluorescence-based assay detection of intracellular calcium mobilization. It is perfect for evaluating GPCR and calcium channel targets as well as for screening their agonists and antagonists. Additionally, Cal-520® AM is completely compatible with use in flow cytometry, fluorescence microscopy, fluorescence spectroscopy and the fluorescence microplate platform. For a sampling of specific applications, please consult the References and Citations section below.
相关产品：
Cal-520® AM is a next-generation fluorescent calcium indicator with an Ex/Em of 492/514 nm. It has several critical advantages and improvements over older generations of fluorescent Ca2+ probes such as Fluo-3 AM and Fluo-4 AM, namely:
- Enhanced Brightness and Sensitivity
- Significantly Higher Signal to Noise Ratio
- More Consistent Loading and Localization
- Convenient Spectrum and Easy Multiplexing
The History
Every single year, there are hundreds of thousands of peer-reviewed papers published focusing on calcium and its role in biological systems. The reason for this is clear. Calcium ions (Ca2+) play a critical part in many biological processes, from the contracting of muscles to the release of neurotransmitters. This pervasiveness of calcium in biology, and the subsequent need to study it, has pushed for the development of viable, convenient calcium indicators, tools that researchers may use to quantify and visualize intracellular calcium.
The first big breakthrough came in the 1980s, when a team of scientists at the University of California Berkeley developed a fluorescein-based calcium indicator. Their research paved the way for now well-established calcium probes such as Fluo-3 AM and Fluo-4 AM. These fluorescein-based AM esters readily pass through the membranes of living cells, where once inside, they are rapidly hydrolyzed by esterases and set to detect calcium. Upon successful binding with calcium, Fluo-3 or Fluo-4 will become fluorescent, emitting a signal at ~520 nm when excited by a ~490 nm light source.
A significant achievement in its own right, these first generation fluorescein-based dyes did however suffer from several major drawbacks. One of these drawbacks was the high background fluorescence, sometimes termed noise. In general, noise creates problems in data analysis and can obscure possibly significant results, and should be minimized as much as possible. That is why we set out to synthesize a new, low noise fluorescent calcium indicator. After much research and optimizing by our dedicated team of experts, we have developed a new high signal to noise ratio calcium indicator, Cal-520® AM.
Cal-520® AM Features
Cal-520® AM is a next-generation fluorescent calcium indicator that has significant improvements over Fluo-3 AM and Fluo-4 AM. First, by improving the binding specificity of Cal-520® for calcium ions, we have managed to produce an indicator which reduces autofluorescence. This simultaneously lowers background fluorescence and raises the signal to noise ratio. The binding of Cal-520® to Ca2+ will result in a greater than one hundred fold increase in fluorescence intensity. Additionally, we have optimized Cal-520®'s chemical structure so that it is better localized. Unlike Rhod-2, which will localize in mitochondria, Cal-520® will predominantly localize in the cytosol. Lastly, we have developed Cal-520® AM for easy use in multiplexing. While Cal-520® AM retains the familiar spectra of Fluo-4 AM and Fluo-8® AM, we also have Cal-590™ AM (Ex/Em = 573/588 nm) and Cal-630™ (Ex/Em = 608/626 nm) for longer wavelength excitations.
Table 1. Various Excitations and Emissions for Multiplex Assays
| Cat No. | Product | Ex (nm) | Em (nm) | Kd (nM) | Additional Notes |
| 21131 | Cal-520™ AM | 492 | 514 | 320 | Best used with 488 nm laser and FITC filter set |
| 20512 | Cal-590™ AM | 573 | 588 | 561 | Best used with 555 nm laser and TRITC/Cy3 filter set |
| 20532 | Cal-630™ AM | 608 | 626 | 792 | Best used with 594 nm laser and Texas Red® filter set |
Because of Cal-520® AM's excellent chemical properties, it is a robust probe for fluorescence-based assay detection of intracellular calcium mobilization. It is perfect for evaluating GPCR and calcium channel targets as well as for screening their agonists and antagonists. Additionally, Cal-520® AM is completely compatible with use in flow cytometry, fluorescence microscopy, fluorescence spectroscopy and the fluorescence microplate platform. For a sampling of specific applications, please consult the References and Citations section below.
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