| Thermo Scientific DyLight 680 Amine-Reactive Dye is an NHS ester-activated derivative of high-performance DyLight 680 used to fluorescently label antibodies and other proteins that are then used as molecular probes for cellular imaging and other fluorescence detection methods.DyLight 680produces near-infrared (IR) fluorescence that replaces other near-IR dyes, including Cy5.5™ dye and Alexa Fluor™ 680, and is ideal for multi-color 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 680 Amine-Reactive Dye is also available as part of two antibody labeling kit sizes.DyLight 680 Amine-Reactive Dye is activated with anN-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 aboutNHS 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 specific amounts of the labeling reagent. This feature allows DyLight 680 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 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 1 mg of IgG or similar quantities of other proteins. The microscale kit contains all of the necessary components to perform five separate labeling reactions using 100 µg of IgG. Both kit sizes include the Amine-Reactive DyLight 680 NHS-ester in convenient single-use vials as well as purification resin and spin columns for the preparation of ready-to-use conjugate. |
| 特点 |
| •High performance—DyLight 680 fluoresces brighter than Alexa Fluor 680 and Cy5.5 dye•Specific—NHS ester-activated dye labels proteins and other molecules at primary amines (-NH2)•Optimized procedure—following the standard protocol results in antibodies with excellent dye:protein ratios and recovery rates for optimum activity and fluorescence labeling |
| 应用 |
| • Primary antibody labeling for immunofluorescence microscopy, immunohistochemistry (IHC), Western blotting, or ELISA assay• Target protein labeling forin vitroandin vivofluorescent detection strategies |
| 数据 |
Immunofluorescence microscopy using DyLight 680 dyeCytokeratin 18 (pseudo-colored yellow) and lamin A (blue) in A549 cells were fluorescently labeled with their respective primary antibodies and DyLight 680-Conjugated Goat Anti-Rabbit or DyLight 405-Conjugated Goat Anti-Mouse Secondary Antibodies, respectively. |
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