DyLight Antibody Labeling Kits

Each Thermo Scientific™ DyLight™ antibody labeling kit contains all the necessary components for three 1-mg labeling reactions of any protein with a molecular weight of 50-150 kDa, and subsequent excess dye removal. The DyLight™ dyes included in these kits are activated with N-hydroxysuccinimide (NHS) esters, which is the most commonly used reactive group for labeling proteins. NHS esters react with primary amines to form stable covalent amide bonds and free NHS groups. The DyLight™ Dyes have absorption spectra ranging from 350 nm to 770 nm (Table 1). These reagents fluoresce across a broad pH range with excitation wavelengths that match the output of most common fluorescence instrumentation. Additionally, the water solubility of the DyLight™ reagents allows conjugations using a high dye-to-protein ratio without precipitation.

Our purification resin and spin columns help eliminate equilibration steps, and reduce the need to collect and monitor gravity-flow fractions. This system helps enable the efficient removal of excess dye and, helping you achieve better protein recovery and a more accurate determination of the dye-to-protein ratio.
Each kit includes:

• DyLight™ Fluor NHS Ester, 3 x 65 μg vials
• Borate Buffer (0.67M), 1 mL 
• Purification Resin, 5 mL 
• Microcentrifuge spin columns, 6 columns 
• Microcentrifuge collection tubes, 12 tubes

The protocol for theDyLight™ Antibody Labeling Kits is simple, consisting of 2 main steps (Figure 1).