Thermo Scientific DyLight 680-4xPEG Amine-Reactive Dye is a derivative of our high-performance DyLight 680 Dye that can be used to fluorescently label antibodies and other proteins.
The DyLight 680-4xPEG dye contains 4 polyethylene glycol (PEG) chains that are non-cytotoxic, enhance fluorescence and reduce nonspecific binding of conjugates made with them. Conjugates made with DyLight 680-4xPEG Dye can be used as molecular probes for cellular imaging and other fluorescence detection methods. The far-red to NIR fluorescence properties of DyLight 680-4xPEG Dye make it especially useful in a variety of biological, chemical, and pharmaceutical applications, including in vivo imaging. The PEG chains also improve solubility of the dyes and labeled molecules in aqueous solution, aid in cell permeability and improve tissue retention.
Highlights:
- High fluorescence intensity – significantly brighter fluorescence than Alexa Fluor 680
- PEGylated – improves solubility in aqueous solution, aids in cell permeability and improves tissue retention in in vivo applications
- Convenient kit sizes – standard and microscale kit sizes to match your experimental needs
Applications:
- Fluorescence microscopy
- In vivo or ex vivo imaging
- Cell-based assays
- Flow cytometry/fluorescence-activated cell sorting (FACS)
Product Details:
DyLight 680-4xPEG Amine-Reactive Dye is activated with an N-hydroxysuccinimide (NHS) ester moiety to react with exposed N-terminal α-amino groups or the ε-amino groups of lysine residues to form stable amide bonds. (Learn more about NHS ester chemistry.)
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| Thermo Scientific DyLight 680-4xPEG Dye is brighter than other NIR fluors. The nuclear envelope of U20S osteosarcoma cells was fluorescently labeled with mouse anti-Lamin A primary antibody and goat anti-mouse secondary antibody conjugated to the indicated dyes. All images were captured using the same exposure time and contrast and brightness settings. |
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Thermo Scientific DyLight 680-4xPEG Dye exhibits rapid biodistribution and tissue clearance. (A). Mice were injected with hydrolyzed DyLight 680-4xPEG Dye via the retro-orbital plexus, and NIR in vivo images were captured pre (Pre)- and post-injection at the indicated time points. Data indicate that the dye is rapidly distributed throughout the mouse body and signicantly cleared between 30m and 1h post-injection. (B) After the final time point, animals were sacrificed and tissues collected for ex vivo imaging to confirm dye accumulation in the kidneys and GI tract, indicative of clearance via the excretory system. Heat map legends indicate signal intensity.
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Typical labeling reactions require the dye to first be dissolved in anhydrous dimethyl formamide (DMF) or another suitable organic solvent before adding a specific molar amount of dye to an amine-free buffer containing the protein to be labeled. However, the high solubility of DyLight Fluors permits protein solutions to be added directly to the labeling reagent. This feature allows DyLight 680-4xPEG Amine-Reactive Dye to be provided in multiple formats with flexible protocols to achieve efficient degrees of labeling.
We also offer Standard and Microscale DyLight 680-4xPEG Antibody Labeling Kits for fast and efficient fluorescent labeling of antibodies. The standard size kit contains all necessary components to perform three separate labeling reactions using 1mg IgG or similar quantities of other proteins. The microscale kit contains all of the necessary components to perform five separate labeling reactions using 100μg IgG. Both kit sizes include the Amine-Reactive DyLight 680-4xPEG NHS-ester in convenient single-use vials as well as purification resin and spin columns for the preparation of ready-to-use conjugate.
 Procedure summary for Thermo Scientific DyLight 680-4xPEG Antibody Labeling Kits. The protocol is simple and provides efficient labeling, exceptional clean-up and excellent antibody recovery. |
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DyLight Antibody Labeling Kits provide outstanding recovery. The percent recovery for the DyLight 680-4xPEG Antibody Labeling Kit is the average of 16 labeling reactions using three different antibodies. The percent recovery for DyLight Microscale Antibody Labeling Kits is the average of 28 labeling reactions using two secondary antibodies and several primary monoclonal antibodies.
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Related Resources:
DyLight 680 Fluorophore Overview Page
Complete DyLight Fluor Product Guide
Related Products:
DyLight 680 Amine-Reactive Dye and Kits (not PEGylated)
DyLight 755 and 800 Dyes and Kits
Far-red and NIR DyLight Specialty Dyes
Dye Removal Columns
Pierce Immunostain Enhancer
Fluorescent Counterstains
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