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

FluorLabeling-fig1
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

Microscale Protein
Labeling Kit

Ex/Em (nm)

Catalog #

Spectra

Comparable dyes

DyLight™ 350

353/432

62276

Description: Excitation and Emission Spectra of DyLight 350 Fluorophore

Alexa Fluor® 350, AMCA

DyLight™ 405

400/420

53021

Description: Excitation and Emission Spectra of DyLight 405 Fluorophore

Alexa Fluor 405, Cascade Blue®

DyLight™ 488

493/518

53025

Description: Excitation and Emission Spectra of DyLight 488 Fluorophore

Alexa Fluor® 488, fluorescein, FITC

DyLight™ 550

562/576

84531

Description: Excitation and Emission Spectra of DyLight 550 Fluorophore

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

DyLight™ 594

593/618

53045

Description: Excitation and Emission Spectra of DyLight 594 Fluorophore

Alexa Fluor® 594, Texas Red®

DyLight™ 633

638/658

53047

Description: Excitation and Emission Spectra of DyLight 633 Fluorophore

Alexa Fluor® 633

DyLight™ 650

652/672

84536

Description: Excitation and emission spectra for DyLight 650

Alexa Fluor 647®, Cy5®

DyLight™ 680

692/712

53057

Description: Excitation and Emission Spectra of DyLight 680 Fluorophore

Alexa Fluor® 680, Cy5.5®

DyLight™ 755

754/776

84539

Description: Excitation and Emission Spectra of DyLight 750 Fluorophore

Alexa Fluor® 750

DyLight™ 800

777/794

53063

Description: Excitation and Emission Spectra of DyLight 800 Fluorophore

IRDye® 800

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Alexa Fluor® Labeling Kits