Multi-Color Fluorescence Microscopy Application using Novel Brilliant Violet™ and Alexa Fluor® 594 Conjugated Monoclonal Antibodies
BioLegend has generated a large library of high affinity antibody clones against different mouse and human antigens that are validated in flow cytometry applications. They are offered as conjugates with the violet-excited Brilliant Violet™ (BV™) fluorophores. Use of the BV™ fluorophores has greatly increased phenotypic multiplexing capabilities in flow cytometry by increasing the utility of the violet laser. Violet-excited BV421™ antibody conjugates are at least as bright as yellow/green-excited phycoerythrin (PE) and red-excited Alexa Fluor® 647 conjugates. The brightness of violet-excited BV510™ is more on par with the green-excited Alexa Fluor® 488. BV421™ and BV510™ are far brighter and more photostable than the violet-excited Alexa Fluor® 405 and Pacific Blue™. Here, using a variety of discrete immunologic cell populations bearing different antigens, BV421™, BV510™, Alexa Fluor® 488, Alexa Fluor® 594 and Alexa Fluor® 647 are shown to also increase multiplexing capabilities and increase resolution in fluorescence microscopy applications. The results show that BV421™, Alexa Fluor® 488, Alexa Fluor® 594 and Alexa Fluor® 647 conjugated monoclonal antibodies are valuable new tools for the immunologist wanting to resolve multiple antigens simultaneously with fluorescence microscopy