Abstract 5674: StarBrightTM Dyes for flow cytometry, western blotting, microscopy and Genesis System with CelSelect Slides

Abstract There are many antibody-based applications in the researcher’s toolbox that rely on both high-quality antibodies and bright stable fluorescent dyes. Antibodies are usually validated for specific applications and may only work on fresh samples and not work if a sample is fixed. The same application specificity can also be observed with fluorescent dyes. Some dyes are easily bleached, may not be stable after fixation and have spectral profiles that make it impossible to multiplex in some applications. To be able to use multiple applications, researchers may need to purchase three or four versions of an antibody or use sub-optimal choices to keep costs manageable. Here we present data on StarBright Dyes, a range of 33 fluorescent dyes, in a multitude of antibody-based applications. They are bright, have low cross laser excitation, can be fixed in PFA and alcohol-based buffers and work in all protocols without specific buffers, especially when multiplexing. StarBright Dyes have been developed to give researchers high quality data whilst providing flexibility and choice regardless of the application. They work in flow cytometry, allowing very high plex panels to be developed with manageable spillover and spreading, require no special buffer and even allow storage of premixed panels. In western blot their brightness, short exposure time and low background makes them ideal for detection of a single target, or in multiplexing. Their brightness allows easy identification and enumeration in cell enrichment systems such as the Genesis SystemTM. Finally in microscopy, both wide field and confocal, their photostability, range of emission, and brightness, not only gives more choice, but as they can be used as primaries without the need for secondary amplification can speed up staining protocols and allow panel expansion without species cross reactivity. Here we demonstrate how you can reduce the number of fluorophores and antibodies required for each application and simplify your protocols by including StarBright Dye conjugated antibodies in your experiments. Citation Format: Michael Blundell, Leon Huang, Sharon Sanderson. StarBrightTM Dyes for flow cytometry, western blotting, microscopy and Genesis System with CelSelect Slides [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 5674.