Thermo Scientific DyLight 550 Amine-Reactive Dye is an NHS ester-activated derivative of high-performance DyLight 550 used to fluorescently label antibodies and other proteins that are then used as molecular probes for cellular imaging and other fluorescence detection methods.
DyLight 550 provides vibrant orange-to-red fluorescence with better performance than other rhodamine derivatives, including Alexa Fluor™ 555, TRITC and Cy3™ dye for fluorescent applications. The high water solubility of DyLight Fluors means that a high dye-to-protein ratio can be attained without causing precipitation of the conjugates. DyLight 550 Amine-Reactive Dye is available as a stand-alone reagent or as part of two antibody labeling kit sizes.
- High performance – DyLight 550 shows brighter fluorescence than Alexa Fluor 555, TRITC and Cy3 dye
- Specific – NHS ester-activated dye labels proteins and other molecules at primary amines (-NH2)
- Convenient kit sizes – standard and microscale sizes are offered to match your experimental needs
- Optimized procedure – following the standard protocol results in antibodies with excellent dye:protein ratios and recovery rates for optimum activity and fluorescence labeling
- Primary antibody labeling for immunofluorescence microscopy, immunohistochemistry (IHC), Western blotting or ELISA assay
- Target protein labeling for in vitro and in vivo fluorescent detection strategies
|Immunofluorescence microscopy using DyLight 550 dye. Cytokeratin 18 (green) and lamin B1(red) in U2OS cells were detected with the respective primary antibodies and goat anti-mouse and anti-rabbit secondary antibodies labeled with DyLight 550 and 650 Microscale Antibody Labeling Kits, respectively.
DyLight 550 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 DyLight 550 Amine-Reactive 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 specific amounts of the labeling reagent. This feature allows DyLight 550 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 550 Antibody Labeling Kits for fast and efficient fluorescent labeling of antibodies for use in fluorescence methods.The standard size kit contains all necessary components to perform three separate labeling reactions using 1mg of IgG or similar quantities of other proteins. The microscale kit contains all of the necessary components to perform five separate labeling reactions using 100µg of IgG. Both kit sizes include the Amine-Reactive DyLight 550 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 DyLight 550 Antibody Labeling Kits. The protocol is simple and provides efficient labeling, exceptional clean-up and excellent antibody recovery.
|DyLight Antibody Labeling Kits provide outstanding recovery. The percent recovery for DyLight Antibody Labeling Kits is the average for 16 labeling reactions using three different antibodies. The percent recovery for DyLight Microscale Antibody Labeling Kits is the average for 13 labeling reactions using three different antibodies.
DyLight 550 Fluorophore Overview Page
Complete DyLight Fluor Product Guide
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