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DyLight 350

Fluorescent dye properties, example data, product guide and references.

Thermo Scientific DyLight 350 is a derivative of AMCA (Aminomethylcoumarine acetate), which is a blue fluorescent dye commonly used as a contrasting dye for double- and triple-labeling used in a wide array of applications. The UV-excitable DyLight 350 has spectral properties that are very similar to those displayed by AMCA, including identical excitation wavelengths and a large stokes shift. The emission wavelength of DyLight 350 has a 10-nm blue shift over AMCA, though, that provides an advantage in multiplexing with green fluorophores. In addition, DyLight 350 has enhanced solubility over AMCA, allowing for higher incorporation of the dye into the biomolecule of interest.

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Fluorescent Probes

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DyLight Fluor Product Guide

DyLight 350 Properties and Applications

DyLight 350 spectral properties
Thermo Scientific DyLight 350 dye spectra. Fluorescent dyes are named based on their excitation (absorption) maxima. The excitation (black) and emission (purple) spectra are normalized to the same height in this graph.

 

Properties of Thermo Scientific DyLight 350.
Parameter Value
Excitation / emission maxima 353nm / 432nm
Emission color Blue
Molar extinction coefficient (ε) 15,000 M-1 cm-1
Correction factor (A280/A353) † 0.144
Molecular weight

NHS ester: 874g/mol
Maleimide: 899g/mol

Spectrally similar dyes Alexa Fluor* 350, AMCA
† Correction factor can be used to estimate protein labeling efficiency.

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Emission spectra of DyLight Fluorescent Dyes.
Compare spectra and properties among all DyLight Fluors

 

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DyLight 800
Fluorescent detection of cytokeratin with DyLight 350
Immunofluorescence microscopy using Thermo Scientific DyLight 350 dye. α-tubulin (blue) and Lamin B1 (green) in A549 cells were fluorescently labeled with their respective primary antibodies and DyLight 350-Conjugated Highly Cross-Adsorbed Goat Anti-Mouse or DyLight 488-Conjugated Goat Anti-Rabbit Secondary Antibodies, respectively.

 

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Fluorescent Probes: Description, Methods and Applications

 

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Thermo Scientific Pierce Fluorescent Products Guide
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DyLight 350 Comparative Data

Cytokeratin detected with AMCA Cytokeratin detected with Alexa Fluor 350 Cytokeratin detected with DyLight 350
Thermo Scientific DyLight 350 brilliant fluorescent staining is substantially brighter than AMCA and evenly comparable to Alexa Fluor* 350. Cytokeratin 18 (blue) was fluorescently labeled in A549 cells with a rabbit polyclonal anti-cytokeratin 18 antibody and goat anti-rabbit IgG secondary antibodies conjugated to AMCA, Alexa Fluor* 350 or DyLight 350. Lamin A (green) was labeled with anti-lamin A antibody and DyLight 488-Conjugated Goat Anti-Mouse Secondary Antibody. All images were captured using identical microscopic and camera settings.

 

DyLight 350 Products

DyLight 350 Reactive Dyes and Kits:

DyLight 350-conjugated Fluorescent Probes:

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DyLight Specialty Dyes

DyLight Fluor Product Guide


Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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