DyLight Microscale Protein 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 microscale protein labeling kit components
Each Thermo Scientific™ DyLight™ microscale protein labeling kit contains all the necessary components for five labeling reactions of 100 μg of protein with molecular weight of 50-150K, 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 over a broad pH range and match the output wavelengths of common fluorescence instrumentation. Additionally, the water solubility of DyLight™ reagents enables 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, 5 x 15 μg vials
- Borate Buffer (0.67M), 1mL
- Purification Resin, 5mL
- Microcentrifuge Spin Columns, 5 each
- Microcentrifuge Collection Tubes, 10 each
DyLight™ microscale protein labeling kit protocol
The protocol for the DyLight™ microscale protein labeling kits is very easy and consists of 2 main steps (Figure 1).
Table 1. DyLight™ microscale protein labeling kit selection guide
Microscale Protein |
Ex/Em (nm) |
Catalog # |
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
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Molecular Probes Handbook
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