Thermo Scientific DyLight microscale protein labeling kits contain NHS ester-activated derivatives of our high-performance DyLight dyes designed specifically for labeling small quantities of antibodies or proteins. Once they are fluorescently labeled, antibodies and other proteins can be used 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 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).
Figure 1. The protocol is simple and provides efficient labeling, exceptional clean-up and excellent antibody recovery.
Table 1. DyLight microscale protein labeling kit selection guide
|Ex/Em (nm)||Cat. No.||Spectra||Comparable dyes|
|DyLight 350||353/432||62276||Alexa Fluor 350, AMCA|
|DyLight 405||400/420||53021||Alexa Fluor 405, Cascade Blue|
|DyLight 488||493/518||53025||Alexa Fluor 488, fluorescein, FITC|
|DyLight 550||562/576||84531||Alexa Fluor 546, Alexa Fluor 555, Cy3,TRITC|
|DyLight 594||593/618||53045||Alexa Fluor 594, Texas Red|
|DyLight 633||638/658||53047||Alexa Fluor 633|
|DyLight 650||652/672||84536||Alexa Fluor 647, Cy5|
|DyLight 680||692/712||53057||Alexa Fluor 680, Cy5.5|
|DyLight 755||754/776||84539||Alexa Fluor 750|
|DyLight 800||777/794||53063||IRDye® 800|
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