Thermo Scientific DyLight 650-4xPEG Amine-Reactive Dye is a derivative of our high-performance DyLight 650 Dye that can be used to fluorescently label antibodies and other proteins for far-red detection.
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.
- High fluorescence intensity – significantly brighter fluorescence than Alexa Fluor™ 647
- PEGylated – improves solubility in aqueous solution and aids in cell permeability
- Fluorescence microscopy
- In vivo or ex vivo imaging
- Cell-based assays
- Flow cytometry/fluorescence-activated cell sorting (FACS)
DyLight 650-4xPEG 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.
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.
|Thermo Scientific DyLight 650-4xPEG Dye is brighter than other spectrally similar fluors. The endoplasmic reticulum in U20S osteosarcoma cells was fluorescently labeled with mouse anti-PDI primary antibody and goat anti-mouse secondary antibody conjugated to DyLight 650-4xPEG, Alexa Fluor 647 (Life Technologies Corp.) or CF 647 (Biotium Inc.) dyes. All images were captured using the same exposure time and contrast and brightness settings.
|Thermo Scientific DyLight 650-4xPEG Dye exhibits rapid biodistribution and tissue clearance. Mice were injected with hydrolyzed DyLight 650-4xPEG or DyLight 650 (non-PEGylated) Dye via the retro-orbital plexus, and NIR in vivo images were captured pre (Pre)- and post-injection at the indicated time points (top images). Data indicate that both dyes are rapidly distributed throughout the mouse body, but the PEGylated dye is more rapidly cleared than the non-PEGylated dye. After the final time point, animals were sacrificed and tissues collected for ex vivo imaging of the kidneys and GI tract to indicate clearance of the dyes via the excretory system. Heat map legends indicate signal intensity (bottom panel).
Click image to enlarge.
|Thermo Scientific DyLight 650-4xPEG Dye is compatible with flow cytometers. The LFA1 cell surface adhesion glycoprotein in Jurkat cells was fluorescently labeled with mouse anti-LFA1 primary antibody and DyLight 650-4xPEG Dye-conjugated goat anti-mouse secondary antibody. The stained cells were then counted using the BD™ Accuri™ C6 Flow Cytometer.
DyLight 650 Fluorophore Overview Page
Complete DyLight Fluor Product Guide
DyLight 650 Amine-Reactive Dye and Kits (not PEGylated)
Dye Removal Columns
Pierce Immunostain Enhancer