_DyLight-Microscale-Protein-Labeling-Kits-230
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 accross 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).

FluorLabeling-fig1
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

Catalog number

Spectra

Comparable Dyes

DyLight™ 350

353/432

62275

Description: Excitation and Emission Spectra of DyLight 350 Fluorophore

Alexa Fluor® 350, AMCA

DyLight™ 405

400/420

53020

Description: Excitation and Emission Spectra of DyLight 405 Fluorophore

Alexa Fluor 405, Cascade Blue®

DyLight™ 488

493/518

53024

Description: Excitation and Emission Spectra of DyLight 488 Fluorophore

Alexa Fluor® 488, fluorescein, FITC

DyLight™ 550

562/576

84530

Description: Excitation and Emission Spectra of DyLight 550 Fluorophore

Alexa Fluor® 546, Alexa Fluor® 555, Cy3®,TRITC

DyLight™ 594

593/618

53044

Description: Excitation and Emission Spectra of DyLight 594 Fluorophore

Alexa Fluor® 594, Texas Red®

DyLight™ 633

638/658

53046

Description: Excitation and Emission Spectra of DyLight 633 Fluorophore

Alexa Fluor® 633

DyLight 650

652/672

84535

Description: Excitation and emission spectra for DyLight 650

Alexa Fluor 647®, Cy5®

DyLight™ 680

692/712

53056

Description: Excitation and Emission Spectra of DyLight 680 Fluorophore

Alexa Fluor® 680, Cy5.5®

DyLight™ 755

754/776

84538

Description: Excitation and Emission Spectra of DyLight 750 Fluorophore

Alexa Fluor® 750

DyLight™ 800

777/794

53062

Description: Excitation and Emission Spectra of DyLight 800 Fluorophore

IRDye® 800

Custom fluorescent antibody and protein labeling service
Alexa Fluor® Labeling Kits