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DyLight 650 Amine-Reactive Dye and Kits


Label amines with a bright alternative to Alexa Fluor™ 647 and Cy5™ dyes.

DyLight 650 Amine-Reactive Dye

Thermo Scientific DyLight 650 Amine-Reactive Dye is an NHS ester-activated derivative of high-performance DyLight 650 used to fluorescently label antibodies and other proteins that are then used as molecular probes for cellular imaging and other fluorescence detection methods.

DyLight 650 provides vibrant far-red fluorescence with comparable or improved performance over other dyes, including Alexa Fluor™ 647 and Cy5™ dye, for fluorescent applications.The high water solubility of DyLight Fluors means that a high dye-to-protein ratio can be attained without causing precipitation of the conjugates. DyLight 650 Amine-Reactive Dye is available as a stand-alone reagent or as part of two antibody labeling kit sizes.

Highlights:

  • High performance – DyLight 650 fluoresces brighter than Alexa Fluor 647 and Cy5
  • Specific – NHS ester-activated dye labels proteins and other molecules at primary amines (-NH2)
  • Convenient kit sizes – standard and microscale sizes are offered to match your experimental needs
  • Optimized procedure – following the standard protocol results in antibodies with excellent dye:protein ratios and recovery rates for optimum activity and fluorescence labeling

Applications:

  • Primary antibody labeling for immunofluorescence microscopy, immunohistochemistry (IHC), Western blotting or ELISA assay
  • Target protein labeling for in vitro and in vivo fluorescent detection strategies

Product Details:

Fluorescent detection of cytokeratin 18 in A549 cells with DyLight 650 by epifluorescence microscopy
Immunofluorescence microscopy using DyLight 650 dye. Intermediate filaments (red) in A549 cells were fluorescently labeled with biotinylated mouse anti-cytokeratin 18 antibody and DyLight 650-Conjugated Streptavidin. Nuclei were labeled with Hoechst dye.

DyLight 650 Amine-Reactive Dye is activated with an N-hydroxysuccinimide (NHS) ester moiety to react with exposed N-terminal α-amino groups or the ε-amino groups of lysine residues to form stable amide bonds. Learn more about NHS ester chemistry.

Typical labeling reactions require the dye to first be dissolved in anhydrous dimethyl formamide (DMF) or another suitable organic solvent before adding a specific molar amount of dye to an amine-free buffer containing the protein to be labeled. However, the high solubility of DyLight Fluors permits protein solutions to be added directly to specific amounts of the labeling reagent. This feature allows DyLight 650 Amine-Reactive Dye to be provided in multiple formats with flexible protocols to achieve efficient degrees of labeling.

We also offer Standard and Microscale DyLight 650 Antibody Labeling Kits for fast and efficient fluorescent labeling of antibodies for use in fluorescence methods.The standard size kit contains all necessary components to perform three separate labeling reactions using 1mg of IgG or similar quantities of other proteins. The microscale kit contains all of the necessary components to perform five separate labeling reactions using 100µg of IgG. Both kit sizes include the Amine-Reactive DyLight 650 NHS-ester in convenient single-use vials as well as purification resin and spin columns for the preparation of ready-to-use conjugate.

Protocol summary for DyLight Antibody Labeling Kits.
Procedure summary for DyLight 650 Antibody Labeling Kits. The protocol is simple and provides efficient labeling, exceptional clean-up and excellent antibody recovery.

DyLight Antibody Labeling Kits provide outstanding recovery.
DyLight Antibody Labeling Kits provide outstanding recovery. The percent recovery for DyLight Antibody Labeling Kits is the average for 16 labeling reactions using three different antibodies. The percent recovery for DyLight Microscale Antibody Labeling Kits is the average for 13 labeling reactions using three different antibodies.

Related Resources:

DyLight 650 Fluorophore Overview Page
Complete DyLight Fluor Product Guide

Related Products:

All Fluorescent Labeling Reagents and Kits
DyLight Fluor Secondary Antibody Conjugates
DyLight Fluor Streptavidin and NeutrAvidin Conjugates
DyLight 650 Azide-Reactive Dye – phosphine-activated fluorescent dye for Staudinger ligation
Krypton Fluorescent Protein Stains


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
62266 DyLight 650 NHS Ester
Formulation: NHS ester activated proprietary far-red fluorescent dye
Sufficient For: 5 uses to label a total of 2mg of IgG using typical conditions

Replaces DyLight Fluor 649
5 x 50µg Product Instructions for product #62266 DyLight 650 NHS Ester MSDS for product #62266 DyLight 650 NHS Ester Certificate of Analysis for product #62266 DyLight 650 NHS Ester $187.00

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62265 DyLight 650 NHS Ester
Formulation: NHS ester activated proprietary far-red fluorescent dye
Sufficient For: 1 use to label a total of 7mg of IgG using typical conditions

Replaces DyLight Fluor 649
1mg Product Instructions for product #62265 DyLight 650 NHS Ester MSDS for product #62265 DyLight 650 NHS Ester Certificate of Analysis for product #62265 DyLight 650 NHS Ester $239.00

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62265B DyLight 650 NHS Ester
Formulation: NHS ester activated proprietary far-red fluorescent dye
Custom MSDS for product #62265B DyLight 650 NHS Ester Certificate of Analysis for product #62265B DyLight 650 NHS Ester
84535 DyLight 650 Antibody Labeling Kit
Formulation: Multi-component kit with proprietary far-red fluorescent dye
Sufficient For: Labeling and purifying 3 x 1mg (2mg/mL) of IgG or other proteins

Kit Contents:
DyLight Fluor NHS Ester, 3 x 50μg vials
Borate Buffer (0.67M), 1mL
Purification Resin, 5mL
Microcentrifuge Spin Columns, 6 columns
Microcentrifuge Collection Tubes, 12 tubes
3-rxn kit Product Instructions for product #84535 DyLight 650 Antibody Labeling Kit MSDS for product #84535 DyLight 650 Antibody Labeling Kit Certificate of Analysis for product #84535 DyLight 650 Antibody Labeling Kit $350.00

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84536 DyLight 650 Microscale Antibody Labeling Kit
Formulation: Multi-component kit with proprietary far-red fluorescent dye
Sufficient For: Labeling and purifying 5 x 100µg (1mg/mL) of IgG or other proteins

Kit Contents:
DyLight Fluor NHS Ester, 5 x 15μg vials
Borate Buffer (0.67M), 1mL
Purification Resin, 5mL
Microcentrifuge Spin Columns, 5 columns
Microcentrifuge Collection Tubes, 10 tubes
5-rxn kit Product Instructions for product #84536 DyLight 650 Microscale Antibody Labeling Kit MSDS for product #84536 DyLight 650 Microscale Antibody Labeling Kit Certificate of Analysis for product #84536 DyLight 650 Microscale Antibody Labeling Kit $328.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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