Successful cell cycle analysis requires a dye that is DNA selective and can stain cells in a homogeneous pattern minimizing fluorescence variability. The Vybrant® DyeCycle™ Ruby Stain is an ideal tool for DNA content analysis in living cells since the stain is cell permeable and, after binding double-stranded DNA, emits a fluorescent signal that is proportional to the DNA mass (see figure).
Low Cytotoxicity
Unlike stains that require high concentrations or have chemical structures that are toxic to cells, Vybrant® DyeCycle™ Ruby Stain exhibits relatively low cytotoxicity, allowing the possibility of sorting based on the phase of the cell cycle.
Minimal Compensation
Well-suited for the popular red 633 nm laser line, the Vybrant® DyeCycle™ Ruby Stain/DNA complex has fluorescence excitation and emission maxima of 638/686 nm, respectively (see figure). The spectral characteristics allow use of the red laser for cell cycle analysis and frees up other channels for additional parameters.
The red excitation and narrow emission of Vybrant® DyeCycle™ Ruby Stain make it ideal for multiplexing due to the limited spectral overlap with other common dyes (Alexa Fluor® 488, FITC, and RPE) and fluorescent proteins (Green Fluorescent Protein (GFP)).
Simple, Robust Staining Protocol
For cell analysis, simply prepare flow cytometry tubes containing 0.5 mL of cells suspended in complete media at a concentration of 5 × 105 cells/mL. To each tube, add 1 µL of Vybrant® DyeCycle™ Ruby Stain and mix well. Final stain concentration is 5 µM. Incubate at 37°C for 15–30 minutes, protected from light. Analyze without washing cells on a flow cytometer using 488 nm or 633/5 nm excitation and >670 nm emission.
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注意事项:
For Research Use Only. Not for use in diagnostic procedures.