AFDye 594 (Alexa Fluor® 594 Maleimide equivalent) is bright, water-soluble, and pH-insensitive from pH 4 to pH 10 red-fluorescent dye with absorption and emission maxima at 590 and 617 nm, respectively. It can be used with the 561 nm and 594 nm laser lines. AFDye 594 dye conjugated to a variety of antibodies, peptides, proteins, tracers, and amplification substrates often used for generation of stable signal in imaging and flow cytometry. The brightness and photostability of this dye are best suited to direct imaging of low-abundance targets.
Maleimide is the most popular sulfhydryl-reactive reactive group for conjugating the dye to a thiol group on a protein, oligonucleotide thiophosphate, or low molecular weight ligand. The maleimide group specifically and efficiently reacts with reduced thiols (sulfhydryl groups, –SH) at pH 6.5 to 7.5 to form stable thioether bond. The resulting conjugates exhibit brighter fluorescence and greater photostability than the conjugates of many other spectrally similar fluorophores.
AFDye 594 dye structurally is identical to Alexa Fluor® 594 C5 Maleimide.