Thermo Scientific DyLight 755 Amine-Reactive Dye is an NHS ester-activated derivative of high-performance DyLight 755 used to fluorescently label antibodies and other proteins that are then used as molecular probes for cellular imaging and other fluorescence detection methods.
DyLight 755 is a near-IR fluor that is invisible to the naked eye but increases the staining options when using infrared imaging systems. DyLight 755 has spectral properties that are very similar to other near-IR dyes, including Alexa Fluor* 750. 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 755 Amine-Reactive Dye is available as a stand-alone reagent or as part of two antibody labeling kit sizes.
Highlights:
- High performance – DyLight 755 replaces Alexa Fluor 755 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:
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Infrared Western blot detection of ERK1 using Thermo Scientific DyLight 755-conjugated secondary antibody. Serial dilutions of A562 cell lysates were analyzed by Western blotting for levels of ERK1 using a mouse anti-ERK1 primary antibody and a goat anti-mouse secondary antibody conjugated to DyLight 755 (using the DyLight 755 Microscale Antibody Labeling Kit). Left lane: Infrared molecular weight marker.
View the physical and spectral properties and comparative data of DyLight 755 Fluorophore.
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DyLight 755 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 755 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 755 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 755 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 755 Antibody Labeling Kits. The protocol is simple and provides efficient labeling, exceptional clean-up and excellent antibody recovery. |
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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.
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