Member since: 2016
Organization: Centro Nacional de Biotecnología
Application Area: To label antibodies and use for immunofluorescence
"It is very easy to use and fast. You can label any primary antibody with DyLight 405."
Thermo Scientific DyLight 405 Amine-Reactive Dye is an NHS ester-activated derivative of high-performance DyLight 405 used to fluorescently label antibodies and other proteins that are then used as molecular probes for cellular imaging and other fluorescence detection methods.
DyLight 405 has a vibrant blue fluorescence intensity that replaces Alexa Fluor™ 405, Cascade™ Blue and AMCA in many 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 405 Amine-Reactive Dye is also available as part of two antibody labeling kit sizes.
Features of the DyLight 405 Amine-Reactive Dye:
• High performance— DyLight 405 shows bright fluorescence and replaces Alexa Fluor 405, Cascade Blue and AMCA
• 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
• 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
DyLight 405 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.
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 405 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 405 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 405 NHS-ester in convenient single-use vials as well as purification resin and spin columns for the preparation of ready-to-use conjugate.