DyLight 800 Amine-Reactive Dye and Kits

Thermo Scientific DyLight 800 Amine-Reactive Dye is an NHS ester-activated derivative of high-performance DyLight 800 used to fluorescently label antibodies and other proteins that are then used as molecular probes for cellular imaging and other fluorescence detection methods.

DyLight 800 is a near-IR fluor that is invisible to the naked eye but increases the staining options when using infrared imaging systems. DyLight 800 has spectral properties that are very similar to other near-IR dyes, including Alexa Fluor* 790 and IRDye* 800. 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 800 Amine-Reactive Dye is available as a stand-alone reagent or as part of two antibody labeling kit sizes.

Highlights:

• High performance – DyLight 800 replaces Alexa Fluor 800 and IRDye 800 for near-infrared staining
• 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

Applications:

• 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

Product Details:

Two-color infrared Western blot detection using Thermo Scientific DyLight 680- and DyLight 800-Conjugated Secondary Antibodies. p53 (53 kDa; red) and cyclophilin B (21 kDa; green) levels in response to siRNA expression in A549 cells were detected by Western blot analysis using Thermo Scientific DyLight 680- (red) and DyLight 800- (green) Conjugated Secondary Antibodies. Beta-catenin (90 kDa, green) was used as the load control. Imaged with the Odyssey* Infrared Imaging System using the 700 and 800 channels.   View the physical and spectral properties and comparative data of DyLight 800 Fluorophore.

DyLight 800 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 specific amounts of the labeling reagent. This feature allows DyLight 800 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 800 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 800 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 800 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.

Related Resources: DyLight 800 Fluorophore Overview Page Complete DyLight Fluor Product Guide Related Products: All DyLight Labeling Reagents and Kits All Fluorescent Labeling Reagents and Kits DyLight Fluor Secondary Antibody Conjugates DyLight Fluor Streptavidin and NeutrAvidin Conjugates DyLight 800 Fluor, free acid – used as controls and calibrants Fluorescent Protein Stains