XFD555 PEG4 DBCO

• Ex/Em: 553/568 nm
• Extinction coefficient: 155,000 cm^-1M^-1
• Reactive Group: DBCO
• Copper-Free Bioorthogonal Labeling: Enables rapid and efficient azide conjugation via copper-free SPAAC
• Bright & Stable: Exhibits intense fluorescence with resilience to photobleaching and pH variations from 4 to 10
• Hydrophilic: Minimizes aggregation, enhancing signal clarity for advanced imaging and live-cell studies

XFD555, manufactured by AAT Bioquest, is structurally identical to Alexa Fluor™ 555 (ThermoFisher). It is a bright orange-fluorescent dye with an excitation optimized for use with either the 488 nm line of the argon-ion laser or the 532 nm line of the frequency-doubled Nd:YAG laser. The high fluorescence quantum yield and high photostability of XFD555 allow for the detection of low-abundance biological structures with great sensitivity. XFD555 demonstrates good aqueous solubility and pH-insensitivity over a broad pH range (pH 4–10), ensuring stable fluorescence generation under varying experimental conditions. XFD555 dye molecules can be attached to proteins at high molar ratios without significant self-quenching, enabling brighter conjugates and more sensitive detection in imaging and flow cytometry.

The DBCO derivative of XFD555 is a highly reactive cycloalkyne optimized for copper-free click chemistry (SPAAC, strain-promoted azide-alkyne cycloaddition). This derivative exhibits a significantly higher reaction rate with azides compared to other cyclooctynes and copper-catalyzed click reactions (CuAAC). Uniquely, DBCO does not react with tetrazines, allowing for its use in bioorthogonal reactions alongside trans-cyclooctenes and tetrazines. For applications where the presence of copper is problematic, XFD555 DBCO serves as an effective alternative to copper-dependent fluorescent alkynes.