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StarBright Violet 515
Excitable by the violet 405 nm laser and emitting at 515 nm, StarBright Violet 515 (SBV515) Dye is a perfect replacement for fluorophores that have emission maxima between 500 and 550 nm, such as Pacific Orange, Krome Orange, vioGreen, and Brilliant Violet 510, which can all be detected with a 525/50 filter (or similar) in your flow cytometry experiments. Minimal excitation by other lasers and a narrow emission profile make StarBright Violet 515 (Figure 1) the perfect dye for inclusion in multicolor panels.
Fig. 1. Excitation and emission spectra for SBV515. The dye is excited maximally by the violet laser (dotted line) with minimal excitation by other lasers. The emission profile is narrow to reduce spillover into neighboring filters.
StarBright Violet 515 Dye is also compatible with spectral flow cytometry and has a unique profile allowing incorporation into new and existing flow cytometry panels. The full spectral profile is shown in Figure 2.
Fig. 2. Dye signature of SBV515. The full emission spectrum of StarBright Violet 515 Dye showing the emission at all wavelengths. Data was collected on the Cytek Aurora Flow Cytometry System using SpectroFlo Software.
StarBright Violet 515 Dye is brighter than other common dyes which emit at similar wavelengths and is up to twice as bright as polymer based dyes conjugated to the same clone (Figure 3).
Combined with the narrow emission to reduce spillover and minimal excitation by other common lasers, StarBright Violet 515 Dye is a great choice for inclusion in multicolor panels.
Fig. 4. StarBright Violet 515 Dye lot-to-lot reproducibility. Human peripheral blood mononuclear cells were stained with 3 lots in triplicate of CD4SBV515 (MCA1267SBV515) (solid lines) and CD4 BV510 (dotted line) for comparison of brightness. Minimal variation between lots of CD4SBV515 is observed. All cells were analyzed on the ZE5 Cell Analyzer.
StarBright Violet 515 Dye has been tested in many common staining buffers with no drop in performance, even after fixation. Furthermore, while they work in special polymer dye staining buffers, there is no requirement to use a special staining buffer, so it can be easily added to existing panels with no change to your experimental protocol.
StarBright Violet 515 Dye is not a tandem dye so is not susceptible to photobleaching, has high lot-to-lot reproducibility (Figure 4), and is stable at 4oC with no loss of signal.
StarBright Violet 515 Dye is compatible with organic fluorophores, protein fluorophores, and polymer fluorophores, including other StarBright Dyes in multicolor panels in common experimental staining protocols and most staining buffers. Examples of staining can be seen in Figure 5.
Fig. 5. Examples of StarBright Violet 515 Dye staining. Human peripheral blood was stained with A, CD3A647 (MCA463A647) and CD27SBV515 (MCA755SBV515) or B, CD45ROPE (MCA461PE) and CD45RASBV515 (MCA88SBV515). All staining was done at 4oC for 30 min in PBS containing 1% BSA after blocking with 10% human serum. Cells were analyzed on the ZE5 Cell Analyzer.
For more information on how the StarBright range can fit into your flow cytometry experiments, please take a look at our Starbright Dyes page.
For more information about fluorophores and immunophenotyping, refer to our flow cytometry resources.
Click on the links below to find out more in-depth information on each topic and view our popular flow cytometry basics guide.