Thermo Scientific DyLight Fluorescent Dyes are a complete family of high-intensity, photostable fluorescent tags for labeling antibodies and other molecular probes. The DyLight Fluors are available as reactive labeling agents and as conjugates of secondary antibodies, biotin-binding proteins and molecular weight markers for use in fluorescence microscopy, flow cytometry, ELISA, high-content screening and other array platforms.

Page Contents:

   • Quick Links to Products
   • Properties of DyLight Fluorophores
   • DyLight Reactive Dyes and Labeling Kits
   • DyLight Conjugated Fluorescent Probes
   • Assay Platforms Based on DyLight Fluors
   • References and Publications

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Quick Links to Products

• Labeling Reagents and Kits
• Conjugates (secondary antibodies)
• Conjugates (biotin-binding streptavidin, etc.)

Properties of DyLight Fluorophores

DyLight Fluors have absorption maxima ranging from 350nm to 777nm, covering the entire visible light spectrum and several key near-infared and infrared wavelengths. Both the absorption and emission properties of the DyLight Fluors match the output (excitation) and detection wavelengths of common fluorescence instrumentation.

The DyLight Dyes exhibit higher fluorescence intensity and photostability than Alexa Fluor*, CyDye* and LI-COR* Dyes in many applications and remain highly fluorescent over a broad pH range (pH 4-9). Additionally, the water solubility of the DyLight Dyes allows a high dye-to-protein ratio to be achieved without causing precipitation of conjugates.

Compare DyLight Dyes and Alexa Fluors for fluorescent intensities of antibody and streptavidin conjugates:
   • DyLight 488 vs. Alexa Fluor 488
   • DyLight 550 vs. Alexa Fluor 555
   • DyLight 650 vs. Alexa Fluor 647
   • DyLight 680 vs. Alexa Fluor 680
   • DyLight 800 vs. IRDye 800

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View absorption and emission spectra of DyLight Fluors

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DyLight Fluorophore
Performance Data

DyLight Reactive Dyes and Labeling Kits

The DyLight Fluorescent Dyes are available in both NHS ester-activated and maleimide-activated forms, which provide for efficient and specific labeling of primary amines and sulfhydryl groups, respectively. Several DyLight Fluors are also available with azide-reactive phosphine groups for detection and labeling using Staudinger ligation chemistry. Certain DyLight Fluors are also available as free acids for use as non-reactive negative controls.

The amine-reactive DyLight Fluors are available as both stand-alone reagents and complete kits optimized for milligram- and microgram-scale antibody or general protein labeling reactions. The DyLight Fluor Antibody Labeling Kits include the amine-reactive DyLight Fluors, buffers and special columns for easy clean-up of the labeling reaction and recovery of the labeled protein. The Dye Removal Columns are also available separately.

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DyLight Fluor Antibody and Protein Labeling Kits

Amine-reactive DyLight Dyes
Sulfhydryl-reactive DyLight Dyes
Azide-reactive DyLight Dyes
Free-acid DyLight Dyes
 

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Thermo Scientific Pierce Fluorescent Products Guide

DyLight Fluorescent Probes (Conjugates)

Pre-conjugated secondary antibodies and biotin-binding proteins of DyLight Fluors make excellent secondary detection reagents for fluorescent assays. Our DyLight Fluor Conjugates is optimally labeled with the highest dye-to-protein ratio (F/P ratio) that does not cause quenching of the fluorescent signal or problems with conjugate solubility.

• Secondary Antibody Conjugates – anti-mouse IgG and anti-rabbit IgG secondary antibodies conjugated to DyLight Fluors make a variety of immunodetection, immunoassay and cellular imaging procedures accessible to fluorescence measurement and multiplexing experimental strategies.
• Highly Cross-Adsorbed Secondary Antibody Conjugates – these forms of secondary antibodies are specially purified to minimize binding to off-target immunoglobulins, making them ideal for multiplex immunofluorescence assays where cross-reactivity is a concern.
• Streptavidin Conjugates – streptavidin biotin-binding protein conjugated to DyLight Fluors enables detection of biotinylated primary antibodies and other biotin-labeled targets for fluorescence methods.
• NeutrAvidin Conjugates – Thermo Scientific NeutrAvidin Biotin-Binding Protein is an economical and equally high-performance alternative to streptavidin for biotin detection methods.

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DyLight Fluor Secondary
Antibody Conjugates

DyLight Fluor Antibodies,
Highly Cross-Adsorbed

DyLight Fluor Streptavidin
Conjugates

DyLight Fluor NeutrAvidin
Protein Conjugates

Assay Platforms Based on DyLight Fluor Conjugates

DyLight Fluor conjugated secondary antibodies and other molecular probes are the basis for a variety of optimized assay platforms for multiplex and high-content analysis of cellular pathway targets.

In-Cell ELISA Kits – The Thermo Scientific Near Infrared In-Cell ELISA Kits use pairs of DyLight 680 and DyLight 800 conjugated secondary antibodies to quantify expression of specific targets in whole cells using an ELISA format. A general kit allows users to develop a target-specific assay using their own primary antibodies. Target-specific kits include optimized primary antibodies for particular proteins of research interest (EGFR, ERK, STAT3, etc.).

High Content Screening Kits – Thermo Scientific Cellomics HCS Reagents Kits are multiplex fluorescent assays for cellular imaging and analysis. Each one of these many kits uses DyLight Fluor Antibodies and other fluorescent probes to detect the cellular expression and translocation of specific proteins and cell structural parameters. When used in combination with appropriate imaging equipment and analysis software, these kits provide extremely powerful strategies for examining cellular responses.

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Near Infrared In-Cell ELISA Kit

Target-specific Near Infrared
In-Cell ELISA Kits

Cellomics High Content
Screening (HCS) Reagent Kits

DyLight Fluor References and Publications

As the use of traditional fluorescein (FITC) and rhodamine (TRITC) probes is replaced by more powerful fluors in immunofluorescent and cellular imaging techniques, more articles are published to demonstrate the versatility and application of DyLight Fluor Technology. We are working to catalog this growing body of literature as a resource for our customers, and we hope to make this available soon. Until then, we offer the following guide to DyLight Fluor posters that have been presented by our researchers.

Poster Presentations Featuring DyLight Products (opens in new page).