Alexa Fluor 555: Properties, Applications, & Dye Info

Can a single dye truly revolutionize the world of scientific imaging? Alexa Fluor 555, a vibrant orange dye, has emerged as a pivotal tool in a wide array of biological applications, offering unparalleled versatility and performance. Its significance lies not only in its unique spectral properties but also in its ease of use and compatibility with various experimental techniques.

The realm of scientific research is constantly evolving, with advancements in technology and techniques shaping the landscape of discovery. Within this dynamic environment, fluorescent dyes play a crucial role, allowing scientists to visualize and analyze biological processes at a microscopic level. Among the myriad of dyes available, Alexa Fluor 555 stands out, owing to its exceptional characteristics and broad applicability.

Developed by Molecular Probes, now a part of Thermo Fisher Scientific under the Invitrogen brand name, Alexa Fluor 555 is a member of a family of fluorescent dyes that have become indispensable tools for researchers worldwide. Its popularity stems from its bright fluorescence, photostability, and ease of conjugation to various biomolecules. These features make it an ideal choice for a wide range of applications, from immunofluorescence microscopy to flow cytometry and beyond.

At its core, Alexa Fluor 555 is a fluorochrome, a molecule that absorbs light at a specific wavelength (excitation peak) and emits light at a longer wavelength (emission peak). In the case of Alexa Fluor 555, the excitation peak is around 555 nm, and the emission peak is around 565 nm. This spectral profile positions it perfectly for use with standard microscopy equipment and allows for clear visualization of labeled targets.

The dye's spectral characteristics make it spectrally similar to other commonly used fluorophores, such as Cy3 (GE Healthcare), TRITC, iFluor 555 (Att Bioquest), and iFluor 560 (Att Bioquest). This spectral similarity allows for the easy integration of Alexa Fluor 555 into existing experimental protocols and enables researchers to leverage existing knowledge and resources. Furthermore, Alexa Fluor 555 conjugated antibodies absorb light maximally around 552 nm and fluoresce with a peak around 572 nm.

One of the key advantages of Alexa Fluor 555 is its versatility. The dye can be conjugated to a wide range of biomolecules, including antibodies, proteins, nucleic acids, and other molecules. This versatility allows researchers to target and visualize a wide range of biological structures and processes. Moreover, conjugates are ready to use in less than twenty minutes, streamlining experimental workflows.

Consider, for instance, the use of Alexa Fluor 555 in immunofluorescence microscopy. In this technique, antibodies specific to a target protein are conjugated to Alexa Fluor 555. When these labeled antibodies are introduced to a sample, they bind to the target protein, and their location can be visualized using a fluorescence microscope. The bright orange fluorescence of Alexa Fluor 555 provides a clear and distinct signal, enabling researchers to identify the location and abundance of the target protein within the cell or tissue.

The dye's utility extends beyond immunofluorescence. It is also widely used in flow cytometry, a technique that allows researchers to analyze individual cells based on their physical and chemical properties. In flow cytometry, cells are labeled with fluorescently conjugated antibodies or dyes, and their fluorescence intensity is measured as they pass through a laser beam. Alexa Fluor 555 is a popular choice for flow cytometry due to its bright fluorescence and compatibility with standard flow cytometers.

Furthermore, Alexa Fluor 555 is frequently employed in applications involving streptavidin and biotin. Streptavidin has a very high binding affinity for biotin, and a conjugate of streptavidin is commonly used together with a conjugate of biotin for specific detection of a variety of proteins, protein motifs, nucleic acids, and other molecules. This system provides a highly sensitive and specific method for detecting and quantifying target molecules.

The dye's exceptional properties are further enhanced by its photostability. Alexa Fluor 555 is less prone to photobleaching than many other fluorescent dyes, meaning that it retains its fluorescence intensity for a longer period of time when exposed to light. This is a crucial advantage in microscopy and other imaging techniques, as it allows researchers to acquire high-quality images without the signal fading quickly.

Alexa Fluor 555 is a bright orange dye, and its vibrant color makes it easily distinguishable from other fluorescent labels. This characteristic is particularly useful in multicolor imaging experiments, where multiple targets are labeled with different fluorescent dyes. The distinct color of Alexa Fluor 555 allows researchers to differentiate between different targets and gain a more comprehensive understanding of the biological system under investigation.

In the context of scientific research, the choice of the right tools can significantly impact the outcome of experiments. Alexa Fluor 555 has become a cornerstone in numerous areas of study, including cell biology, immunology, and molecular biology. Its ability to provide high-quality, easily interpretable data makes it an invaluable asset for researchers. It finds its application in the following areas:

• Immunofluorescence Microscopy: Used to visualize the location and abundance of proteins and other molecules within cells and tissues.
• Flow Cytometry: Employed to analyze individual cells based on their physical and chemical properties.
• Protein Detection: Used in techniques like Western blotting and ELISA to detect and quantify proteins.
• Nucleic Acid Detection: Used in techniques like FISH and PCR to visualize and analyze DNA and RNA.
• High-Throughput Screening: Utilized in drug discovery and other screening applications.

The adaptability of Alexa Fluor 555 is not just limited to established techniques. Its chemical properties also make it an excellent choice for innovative research areas. Its use in super-resolution microscopy, for example, allows scientists to exceed the diffraction limit and observe structures at even finer resolutions.

However, it is important to note that this product is intended for use on fixed and permeabilized samples due to the toxicity associated with phalloidin, as in the case of Figure 1. a muntjac skin fibroblast was fixed and permeabilized. While the dye itself is generally safe, the experimental conditions and the materials used in conjunction with it should be considered. Researchers are always advised to follow established safety protocols and guidelines when working with fluorescent dyes.

In summary, Alexa Fluor 555 is a remarkably versatile and valuable tool in the world of scientific research. Its bright fluorescence, photostability, and ease of use make it an ideal choice for a wide range of applications. Whether used in immunofluorescence microscopy, flow cytometry, or other imaging techniques, Alexa Fluor 555 provides researchers with a powerful means of visualizing and analyzing biological processes. The dye's continued development and use are contributing to groundbreaking discoveries in the life sciences, propelling our understanding of the intricate workings of the biological world.

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It has a role as a fluorochrome.

Other spectra of interest include:

Alexa Fluor 555 belongs to the following categories:

X 21422 alexa fluor 555 dyea superior alternative to tetramethylrhodamine and cy3 product information revised:

Furthermore conjugates are ready to use in less than twenty minutes.

Characteristic	Details
Name	Alexa Fluor 555 (AF555, Alexa 555)
Type	Fluorescent Dye
Excitation Peak	555 nm (varies slightly depending on conjugation)
Emission Peak	565 nm (varies slightly depending on conjugation)
Color	Bright Orange
Spectral Similarity	Cy3 (GE Healthcare), TRITC, iFluor 555 (Att Bioquest), iFluor 560 (Att Bioquest)
Applications	Immunofluorescence, Flow Cytometry, Protein Detection, Nucleic Acid Detection, High-Throughput Screening, Super-Resolution Microscopy
Key Features	Bright Fluorescence, Photostability, Ease of Conjugation, Versatility
Developed By	Molecular Probes (Invitrogen, Thermo Fisher Scientific)
Conjugation Time	Ready to use in less than 20 minutes
Used with	Conjugates are commonly used together with a conjugate of biotin for specific detection of a variety of proteins, protein motifs, nucleic acids, and other molecules
Target Samples	Fixed and permeabilized samples

For further information, visit: Invitrogen (Thermo Fisher Scientific)

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