Thermo Scientific DyLight antibody labeling kits contain NHS ester-activated derivatives of our high-performance DyLight dyes. These kits are suitable for moderate scale labeling activities to fluorescently label antibodies and other proteins for use as probes for cellular imaging and other fluorescence detection methods.
- High performance–DyLight dyes show bright fluorescence compared to classic dyes like FITC and rhodamine.
- Specific–NHS ester-activated dye labels proteins and other molecules at primary amines (-NH2)
- Optimized procedure–following the standard protocol results in antibodies with dye:protein ratios and recovery rates that help optimize fluorescence labeling and activity
DyLight antibody labeling kit components
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
DyLight Antibody Labeling Kit Protocol
The protocol for the DyLight Antibody Labeling Kits is simple, consisting of 2 main steps (Figure 1).
Figure 1. The protocol is simple and provides efficient labeling, exceptional clean-up and excellent antibody recovery.
Table 1. DyLight antibody labeling kit selection guide
|DyLight Antibody Labeling Kit||Ex/Em||Cat. No.||Spectra||Comparable Dyes|
|DyLight 350||353/432||62275||Alexa Fluor 350, AMCA|
|DyLight 405||400/420||53020||Alexa Fluor 405, Cascade Blue|
|DyLight 488||493/518||53024||Alexa Fluor 488, fluorescein, FITC|
|DyLight 550||562/576||84530||Alexa Fluor 546, Alexa Fluor555, Cy3,TRITC|
|DyLight 594||593/618||53044||Alexa Fluor 594, Texas Red|
|DyLight 633||638/658||53046||Alexa Fluor 633|
|DyLight 650||652/672||84535||Alexa Fluor 647,Cy5|
|DyLight 680||692/712||53056||Alexa Fluor 680, Cy5.5|
|DyLight 755||754/776||84538||Alexa Fluor 750|
|DyLight 800||777/794||53062||IRDye® 800|
BioProbes Journal articles
Molecular Probes Handbook
For Research Use Only. Not for use in diagnostic procedures.