Label peptides, antibodies and other proteins at primary amines with rhodamine- and benzopyrillium-based Thermo Scientific DyLight Orange-emitting Specialty Dyes, which are comparable to Alexa Fluor 594, Alexa Fluor 610, Cy3.5 and Texas Red.
DyLight Orange-emitting Dyes are a family of labeling agents that provide bright fluorescent detection for imaging. Dyes can be selected based upon characteristic excitation and emission properties or relative hydrophilicity and hydrophobicity attributes. Dyes that contain a greater number of negatively charged sulfonates generally will have greater water solubility than dyes with fewer sulfonates. More hydrophobic dyes often provide better cell penetrating ability in vivo, while more hydrophilic dyes have less nonspecific binding potential. Each dye contains an amine-reactive NHS ester for simple modification of antibodies, proteins, peptides or other biomolecules through amide bond formation.
- NHS ester reactive group – allows immediate labeling of antibodies, proteins, peptides and other amine-containing molecules through amide bond formation
- Multiple solubility options – choose from hydrophilic to hydrophobic dyes to optimize the right dye label for the best performance in a given application
Criteria to consider when choosing a DyLight Orange-emitting Specialty Dye:
- Excitation and emission wavelengths – choose the best dye to match the excitation and emission capabilities of your instrument
- Water solubility – choose a dye based on its relative hydrophilicity, which directly correlates to the number of negatively-charged sulfonates it has on its core structure. More hydrophilic dyes are best at maintaining water solubility of a labeled antibody and limiting the nonspecific binding of the conjugate. More hydrophobic dyes often are best at penetrating tissues and cell membranes in vivo, meaning that dyes with fewer sulfonates may work best for some applications.
- Antibody labeling
- Direct immunofluorescence staining
- Flow cytometry
- Fluorescence correlation spectroscopy
- Western blotting
- Protein microarrays
- Polymer labeling
- Peptide labeling
- Phalloidin labeling for actin staining
- Staining in acidic media
- Biofilm microorganism staining
Thermo Scientific DyLight Orange-emitting Specialty Dye Characteristics.
||Laser and filter sets
||Rhodamine, 2 sulfonates
||546 laser source
532 flow cytometry laser line
Cy3 filter set
546 mercury arc emission
||Benzopyrillium, 1 sulfonate
|† Fluorescent dye name appended with abbreviated core structure (e.g., B2: Benzopyrillium with 2 sulfonates)
†† Molar extinction coefficient (M-1 cm-1) at the absorption maximum
- Brockmann S. et al. (2010) Visualizing acidophilic microorganisms in biofilm communities using acid stable fluorescence dyes. J Fluoresc. 20, 943-51.
- Mindt T. L. et al. (2008) Modification of different IgG1 antibodies via glutamine and lysine using bacterial and human tissue transglutaminase. Bioconjug Chem. 19, 271-8.
- Schroder C. et al. (2010) Dual-color proteomic profiling of complex samples with a microarray of 810 cancer-related antibodies. Mol Cell Proteomics. 9, 1271-80.
- Hu S. et al. (2009) Biotin induced fluorescence enhancement in resonance energy transfer and application for bioassay. Talanta. 80, 454-8.
Fluorescent labeling technical guide
NHS ester crosslinking chemistry technical guide
DyLight 641Q-NHS Ester and 662Q-NHS Ester (complementary fluorescence quenchers)
DyLight 594 Amine-Reactive Dye and Kits
TRITC and Rhodamine Dyes and Labeling Kits