Update profile
    Change password
    Sign out

Phosphine-activated DyLight Fluors


Fluorescent probes to detect azide-tagged cellular targets by Staudinger ligation.

DyLight Fluor Phosphine Reagents

Thermo Scientific DyLight Fluor Phosphine Reagents are phosphine-activated fluorescent dyes for specific labeling and detection of azide-tagged molecules, which enables use of fluorescence imaging in metabolic labeling strategies.

When used in combination with azide labeling strategies, phosphine-activated DyLight Fluors enable selective fluorescent labeling for detection of protein interactions and post-translational modifications using fluorescence imaging technologies. The phosphine group conjugates to azide groups by the Staudinger reaction mechanism. Azide groups can be introduced into proteins or other cellular targets through in vivo labeling with azide-tagged derivatives of naturally occurring metabolic building blocks. Because neither phosphines nor azides are present in biological systems, they comprise a chemoselective (mutually specific) ligation pair for labeling and conjugation.

Highlights:

  • Soluble – easily dissolves in water-miscible solvents (e.g., DMSO) for subsequent dilution in aqueous reaction mixtures with cell lysates and other biological samples
  • Compatible – reaction chemistry occurs effectively in simple buffer conditions; requires no accessory reagents such as copper or reducing agents, and does not interfere with fluorescence applications
  • Chemoselective – the phosphine reactive group is specific in biological samples for bioorthogonal azide-tagged molecules, ensuring that fluorescent labeling is specific
  • High-performance fluorescence – DyLight 488, 550 and 650 are intense, highly stable fluorophores for green, orange and red fluorescent detection. (see DyLight Fluors)

Product Details:

Overview of Staudinger ligation for biomolecular conjugation.
Overview of Staudinger ligation (azide-phosphine conjugation). Phosphine-activated compounds conjugate with high specificity to azide-tagged molecules, resulting in stable covalent attachment of "A" and "B" molecules. For additional information, see our review of Staudinger ligation reaction chemistry.

Overview of Staudinger ligation for biomolecular conjugation.
In vivo detection of metabolically incorporated azido-acetylgalactosamine using Thermo Scientific DyLight 550- and 650-Phosphine Labeling Reagents. Panel A. U2OS cells were incubated with 40µM azido-acetylgalactosamine in cell culture media for 72 hours and the live cells were incubated with 100µM of DyLight 550-phosphine . The cells were then washed, fixed with 4% paraformaldehyde and counterstained with Hoechst 33342. (green: DyLight 550-labeled azido-galactosamine, blue: Hoechst 33342 labeled nuclei). Panel B. HK-2 cells were incubated with 40µM azido-acetylmannosamine in cell culture media for 72 hours and the live cells were incubated with 100µM of DyLight 650-phosphine. The cells were then washed, fixed with 4% paraformaldehyde and counterstained with Hoechst 33342. (cyan: DyLight 650-labeled azido-mannosamine, red: Hoechst 33342 labeled nuclei).

Overview of Staudinger ligation for biomolecular conjugation.
Detection of metabolically incorporated azido-sugars on live and fixed cells using Thermo Scientific DyLight DyLight 550- and 650-Phosphine Labeling Reagents. HK-2 cells were incubated with 40µM azido-acetylmannosamine in cell culture media for 72 hours. The azido-sugar were labeled either after 4% paraformaldehyde fixation using DyLight 550-phoshine (Panel A) or labeled in live cells with DyLight 650-Phosphine Labeling Reagent (Panel B). The cells were washed and counterstained with Hoechst 33342. Panel A. The golgi structure (green) is predominantly detected by fixed cell labeling, where as the cell membrane and secretory vesicles (red) are labeled by live-cell labeling (blue: Hoechst 33342 labeled nuclei).

Related Resources:

Azide-Phosphine Reagent Guide
Review of Metabolic Labeling for Chemoselective Ligation
Review of Staudinger Ligation Reaction Chemistry
Superiority of Azide-Phosphine Chemistry

Related Products:

Azide-Sugars for Metabolic Labeling and Detection
NHS-Azide Staudinger Ligation Reagents
NHS-Phosphine Staudinger Ligation Reagents
Biotin-PEG3-Phosphine


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
88907 DyLight 488-Phosphine
Formulation: Phosphine-activated proprietary green fluorescent dye
Sufficient For: Labeling approx. 5mg of azide-labeled protein using typical conditions
1mg Product Instructions for product #88907 DyLight 488-Phosphine MSDS for product #88907 DyLight 488-Phosphine $362.00

Add to cart
88910 DyLight 550-Phosphine
Formulation: Phosphine-activated proprietary orange fluorescent dye
Sufficient For: Labeling approx. 5mg of azide-labeled protein using typical conditions
1mg Product Instructions for product #88910 DyLight 550-Phosphine MSDS for product #88910 DyLight 550-Phosphine Certificate of Analysis for product #88910 DyLight 550-Phosphine $351.00

Add to cart
88911 DyLight 650-Phosphine
Formulation: Phosphine-activated proprietary far-red fluorescent dye
Sufficient For: Labeling approx. 5mg of azide-labeled protein using typical conditions
1mg Product Instructions for product #88911 DyLight 650-Phosphine MSDS for product #88911 DyLight 650-Phosphine Certificate of Analysis for product #88911 DyLight 650-Phosphine $351.00

Add to cart

Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

Share This Page
 
 

Follow Us
  Email Sign-up  Email Announcements

 



Antibodies  |   Molecular Biology   |   Cell Biology   |  Thermo Scientific  |   * Trademarks  |   Privacy Statement
© 2014 Thermo Fisher Scientific Inc.   |   3747 N Meridian Rd, Rockford, IL USA 61101   |   1-800-874-3723  or  815-968-0747