Expanding multiplexing capabilities with TrailBlazer StarBright Dye antibody conjugation kits

Imaging, western blotting, and flow cytometry multiplexing experiments are often constrained by the availability of suitable fluorophores for some antibodies, hindering panel size and optimal experimental design. Here we address those issues using TrailBlazer™ StarBright™ Dye Antibody Labeling Kits. Custom antibody-dye conjugates can be generated using labeling kits, however options are typically restricted to standard fluorophores, such as FITC, which can be suboptimal and limit panel expansion. The newly introduced TrailBlazer Tag and Label Kits have been launched to enable antibody conjugation to superior StarBright Dyes, with 32 options spanning the five main laser lines. Our data shows that antibodies conjugated with TrailBlazer Kits are stable for up to one year, owing to the novel SpyTag-SpyCatcher technology used. This makes them ideal for long-term storage and eliminates the need for repeated labeling reactions, giving reproducible data particularly in longitudinal studies. We also present new data illustrating the versatility of these conjugates in various applications. This includes immunocytochemical (ICC) staining microscopy data, where they provide additional options when multiplexing by using kit-conjugated primary or secondary antibodies. In addition, flow cytometry data show that panels can be expanded to include antibodies that have limited fluorophore options, without compromising optimal panel design and without the use of secondary antibodies. In summary, TrailBlazer Kits offer a significant advance for researchers, delivering an easy self-conjugation method and enabling the generation of robust, reproducible multiplexing data in complex biological studies in multiple applications.