Why use DyLight® Fluorophores?
DyLight® Fluorescent Dyes are high-intensity, photo-stable fluorescent tags Leinco Technologies uses to label its antibodies and secondary reagents. The DyLight® Fluorescents are available for use in fluorescence microscopy, flow cytometry, Western blotting, ELISA, high-content screening and other array platforms.
For years, researchers have used dyes like fluorescein, rhodamine, Cy3 and Cy5 in a wide variety of applications. However, these dyes have limitations (such as photobleaching) for use in microscopy and flow cytometry which both require exposure to an intense light source. Certain characteristics of the DyLights® make them a highly effective alternative to these dyes in many applications.
- Excitation and emission properties match output
and detection wavelengths of common instrumentation
- Higher photostability / Less pH-sensitive
- Little spectral overlap of wavelengths
Excitation and Emission Properties
The DyLight® Fluorophores are spectrally similar to other dyes, and their excitation and emission properties match the output and detection wavelengths of most fluorescence instrumentation. The absorption spectra of DyLights® carried by Leinco Technologies range from 400–646 nm, covering the entire visible light spectrum and several key near-infrared and infrared wavelengths
Excitation (left) and emission (right) spectra of DyLight® fluorescent dyes conjugated to affinity-purified secondary antibodies. This figure illustrates the relative shape and position of each fluorophore in the peak area of its excitation and emission after antibody conjugation.
|Conjugate||Color||Absorb (nm)||Emit (nm)||ε (M-1cm-1)||QY %||Brightness= (QY*e)/100||Excitation laser||Application Compatibility|
|DyLight® 650||Red||654||673||250,000||31.6||79,000||Red (633 nm)||IF, FC, IHC, ELISA|
|DyLight® 594||Orange||593||618||80,000||80.6||64,480||Green (526 nm)||IF, FC, IHC, ELISA|
|DyLight® 488||Green||493||518||70,000||89.6||62,720||Cyan (488 nm)||IF, FC, IHC, ELISA|
|DyLight® 755||Near-IR||754||776||220,000||26.1||57,420||NIR||WB, ELISA, FC|
|DyLight® 405||Violet||400||420||30,000||96.8||29,040||Laser Diode (405 nm)||IF, FC|
|DyLight® 550||Yellow||562||576||150,000||9.9||14,850||Green (526 nm)||IF, FC, WB, ELISA|
The Dylight® fluorophores show a higher fluorescence strength than comparable dyes like Alexa Fluor, CyDye and LI-COR in most applications. They also maintain greater photostability and are less pH-sensitive. These characteristics allow more time for image capturing.
DyLights® are brighter than Alexa Fluor dyes.
Low Spectral Overlap
Another advantage of the DyLight® Fluorophores is the low spectral overlap of wavelengths. By using a combination of these dyes, it is possible to achieve highly effective 4-color imaging with a maximum separation of colors.
DyLight® is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.