DyLight Antibody Labeling Kits
- 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 theDyLight™ Antibody Labeling Kits is simple, consisting of 2 main steps (Figure 1).
DyLight™ Antibody Labeling Kit |
Ex/Em |
Catalog number |
Spectra |
Comparable Dyes |
---|---|---|---|---|
DyLight™ 350 |
353/432 |
|
Alexa Fluor® 350, AMCA |
|
DyLight™ 405 |
400/420 |
|
Alexa Fluor 405, Cascade Blue® |
|
DyLight™ 488 |
493/518 |
|
Alexa Fluor® 488, fluorescein, FITC |
|
DyLight™ 550 |
562/576 |
|
||
DyLight™ 594 |
593/618 |
|
||
DyLight™ 633 |
638/658 |
|
Alexa Fluor® 633 |
|
DyLight 650 |
652/672 |
|
||
DyLight™ 680 |
692/712 |
|
Alexa Fluor® 680, Cy5.5® |
|
DyLight™ 755 |
754/776 |
|
||
DyLight™ 800 |
777/794 |
|
IRDye® 800 |
Research Tools
BioProbes Journal articles
Molecular Probes Handbook
For Research Use Only. Not for use in diagnostic procedures.