Multi-color fluorescence microscopy detection of chemokine receptors using novel Brilliant Violet™-conjugated monoclonal antibodies (CCR5P.267)

Abstract BioLegend has generated a large library of high affinity antibody clones against mouse and human chemokine receptors that are validated in flow cytometry applications. They are offered as conjugates with the violet-excited Brilliant VioletTM (BVTM) fluorophores. Use of the BVTM fluorophores has greatly increased phenotypic multiplexing capabilities in flow cytometry by increasing the utility of the violet laser. Violet-excited BV421TM 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 BV510TM is more on par with the green-excited Alexa Fluor® 488. BV421TM and BV510TM are far brighter and more photostable than the violet-excited Alexa Fluor® 405 and Pacific BlueTM. Here, using a variety of discrete immunologic cell populations bearing different chemokine receptors, BV421TM and BV510TM are shown to also increase multiplexing capabilities and increase resolution in fluorescence microscopy applications. Elsewhere, both BV421TM and BV510TM were excited with a multiphoton laser, indicating their potential utility in deep tissue and live cell imaging applications. The results show that BV421TM- and BV510TM-conjugated monoclonal antibodies are valuable new tools for the scientist wanting to resolve multiple antigens simultaneously with fluorescence microscopy.