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Alexa Fluor and Alexa Fluor Plus Dyes |
Invitrogen Alexa Fluor and Alexa Fluor Plus dyes are widely used and trusted fluorescent dyes. These dyes are spectrally diverse, high-performing, small molecule fluorophores that are compatible with a variety of fluorescence-based applications. In addition to excellent performance, you'll benefit from an experienced, problem-solving technical support team, over 80,000 published references, and a wealth of application and experimental tips and protocols to aid in experimental planning.
Summary of Alexa Fluor and Alexa Fluor Plus dyes
With the wide range of Invitrogen Alexa Fluor and Alexa Fluor Plus dyes there is something for every filter and many options for multiplex detection. This series of fluorophores spans near-UV, visible and near-IR spectrum and are compatible with all common excitation sources and instruments.
| Color | Alexa Fluor and Alexa Fluor Plus dyes | Excitation/Emission | Dye equivalents |
|---|---|---|---|
| Blue | Alexa Fluor 350 | 343/441 | AMCA, coumarin |
| Alexa Fluor 405 | 401/422 | ||
| Alexa Fluor Plus 405 | 404/455 | ||
| Green | Alexa Fluor 430 | 431/540 | |
| Alexa Fluor 488 | 499/520 | Cy2, FITC (fluorescein) | |
| Alexa Fluor Plus 488 | 493/518 | ||
| Alexa Fluor 514 | 518/543 | ||
| Yellow | Alexa Fluor 532 | 534/553 | |
| Orange | Alexa Fluor 546 | 561/572 | |
| Alexa Fluor 555 | 553/568 | Cy3, TRITC (rhodamine) | |
| Alexa Fluor Plus 555 | 558/572 | ||
| Alexa Fluor 561 | 556/569 | ||
| Red | Alexa Fluor 568 | 579/603 | Rhodamine red |
| Alexa Fluor 594 | 590/618 | Texas Red | |
| Alexa Fluor Plus 594 | 590/618 | ||
| Far Red | Alexa Fluor 633 | 631/650 | |
| Alexa Fluor 635 | 633/648 | ||
| Alexa Fluor 647 | 650/671 | Cy5 | |
| Alexa Fluor Plus 647 | 658/675 | ||
| Near-IR | Alexa Fluor 660 | 663/691 | |
| Alexa Fluor 680 | 681/704 | Cy5.5, IR680 | |
| Alexa Fluor Plus 680 | 687/704 | ||
| Alexa Fluor 700 | 696/719 | ||
| Alexa Fluor 750 | 752/776 | Cy7 | |
| Alexa Fluor 790 | 782/805 | ||
| Alexa Fluor Plus 800 | 789/794 |
Alexa Fluor and Alexa Fluor Plus products
Fluorescent streptavidin & avidin
Protein labeling reagents
Alexa Fluor and Alexa Fluor Plus resources
Advantages of Alexa Fluor dyes
Exceptional brightness
Protein conjugates made with Alexa Fluor dyes produce fluorescence output that surpasses that of other spectrally similar fluorophore-labeled proteins. Alexa Fluor dyes are available in a wide selection of fluorescent colors and are more photostable than most other dyes.
Alexa Fluor dye molecules can be attached to proteins without significant self-quenching, leading to brighter conjugates available for imaging, flow cytometry, and other fluorescence-based applications.
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Comparison of relative fluorescence of goat anti–mouse IgG antibody conjugates prepared from the Alexa Fluor 488 dye and fluorescein isothiocyanate (FITC). Conjugate fluorescence is measured by comparing the fluorescence quantum yield of the conjugated dye relative to that of a reference dye and multiplying by the dye:protein labeling ratio.
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Comparison of relative fluorescence of goat anti–rabbit IgG antibody conjugates of the Alexa Fluor 555 and Cy3 dyes at different dye:protein ratios in the conjugate.
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Flow cytometry comparison of the brightness of Alexa Fluor 647 goat anti–mouse IgG antibody (red, A21235) with Cy5 goat anti–mouse IgG antibody from Jackson ImmunoResearch Laboratories (green) and Amersham-Pharmacia Biotech (blue). Human blood was blocked with normal goat serum and incubated with an anti-CD3 mouse monoclonal antibody; cells were washed, resuspended and incubated with either an Alexa Fluor 647 or Cy5 goat anti–mouse IgG secondary antibody. Samples were analyzed on a flow cytometer equipped with a 633 nm He–Ne laser and a longpass emission filter (>650 nm).
Improved photostability
Photostability is a key characteristic of Alexa Fluor dye conjugates, allowing more time for image capture. In addition to their exceptional brightness and the wide selection of dyes across the spectrum, you'll get photostability you can rely on with Alexa Fluor dyes and dye conjugates.
The photostability of Alexa Fluor dye molecules can be seen when compared to traditional fluorophores such as fluorescein.
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Comparison of the photobleaching rates of the Alexa Fluor 488 and Alexa Fluor 546 dyes and the well-known fluorescein and Cy3 fluorophores. The cytoskeleton of bovine pulmonary artery endothelial cells (BPAEC) was labeled with (top series) Alexa Fluor 488 phalloidin and mouse monoclonal anti–a-tubulin antibody in combination with Alexa Fluor 546 goat anti–mouse IgG antibody or (bottom series) fluorescein phalloidin and the anti–a-tubulin antibody in combination with a commercially available Cy3 goat anti–mouse IgG antibody. The pseudocolored images were taken at 30-second intervals (0, 30, 90, and 210 seconds of exposure). The images were acquired with bandpass filter sets appropriate for fluorescein and rhodamine.
Reduced photobleaching, sustained signal. Bovine pulmonary artery endothelial cells were labeled with fluorescein phalloidin (A) or Alexa Fluor 488 phalloidin (B) which label filamentous actin. The cells were placed under constant illumination on the microscope. Images were acquired at one-second intervals for 30 seconds. Under these illumination conditions, fluorescein images are photobleached to about 20% of its initial value, while fluorescence of Alexa Fluor 488 phalloidin stayed at the initial value.
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Additional advantages of Alexa Fluor Plus dyes
Detection of low-abundance targets with Alexa Fluor Plus dyes
While traditional Alexa Fluor dyes can be used in a variety of applications to detection proteins of varying expression levels, Alexa Fluor Plus dyes have been chemically optimized to provide higher signal to noise ratios and minimized cross-reactivity to help detect low-abundance targets in rare or precious samples.
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Alexa Fluor Plus 405 conjugates allow for improved signal to noise when compared to traditional Alexa Fluor 405 conjugates. Traditional Alexa Fluor 405 secondary antibody (left) compared to Alexa Fluor Plus 405 secondary antibody (right, goat anti-rabbit IgG secondary antibody, Cat. No. A48254) in immunofluorescent analysis of β-III tubulin in fixed HeLa cells. Nuclei were stained with SYTOX Deep Red stain (Cat. No S11380). The images contain overlay of tubulin (cyan), and nuclei (magenta). Images were taken on an EVOS M7000 Imaging system (Cat. No. AMF7000) at 20x magnification.
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Alexa Fluor Plus 555 allows you to see greater detail compared to traditional Alexa Fluor 555 conjugates. Traditional Alexa Fluor 555 secondary antibody (left) compared to Alexa Fluor Plus 555 secondary antibody (right, goat anti-mouse IgG secondary antibody, Cat. No. A32727) in immunofluorescent analysis of β-III tubulin in E18 Sprague Dawley primary cortical neuronal cells. Nuclei were stained with Hoechst 33342 stain (Cat. No. H3570). The image contains overlay of neuronal tubulin (orange) and nuclei (blue). Images were taken on a confocal microscope at 40x magnification.
For Research Use Only. Not for use in diagnostic procedures.
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Not for resale. Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
Brilliant Ultra Violet™ and Brilliant Violet™ are trademarks or registered trademarks of Becton, Dickinson and Company or its affiliates, and is used under license.
For Research Use Only. Not for use in diagnostic procedures.