XFD555, manufactured by AAT Bioquest, is structurally similar to Alexa Fluor™ 555 (Thermo Fisher). 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 N-hydroxysuccinimidyl (NHS) ester of XFD555 is a widely used reagent for the conjugation of this dye to proteins or antibodies. NHS esters react selectively and efficiently with primary amines (such as the side chains of lysine residues or aminosilane-coated surfaces) at pH 7–9, forming stable covalent amide bonds. This property makes XFD555 NHS ester an excellent choice for labeling proteins, amine-modified oligonucleotides, and other amine-containing molecules.