BODIPY 630/650X

BODIPY 630/650X is a red fluorescent dye that targets amine groups. BODIPY 630/650X displays excitation/emission maxima of 630/650 nm, respectively. BODIPY 630/650X can be used to covalently label proteins, antibodies, or other amine-containing biomolecules for detection and imaging^[1]. In Vitro:BODIPY 630/650X is beneficial when imaging in living cells as it is clearly distinguishable from background cell autofluorescence emission^[1].
Live-cell Imaging Protocol^[2][3]:
1.Dye Stock Solution Preparation: Prepare a stock solution of BODIPY 630/650-X at a concentration of 1 mg/mL in DMSO and kept at 4°C in the dark.
2.Working Solution Preparation: Dilut BODIPY 630/650X in PBS to obtain a final working concentration of 5 µg/mL. Prepare the working solution fresh immediately before use and protect from light until use.
3.Cell Staining:
(1)Wash cells twice with pre-warmed PBS to remove serum components.
(2)Incubate the cells with 5 μg/mL BODIPY 630/650X for 25-30 min at room temperature.
4.Washing: Wash cells 3-4 times with complete culture medium (or PBS) containing 5-10% serum or 1% BSA to quench unbound dye. Perform a final wash with live cell imaging buffer.
5.Imaging: Acquire images immediately using a confocal microscope or high content imaging system. Excited with a HeNe laser (wavelength 633 nm) and collect emission using a long-pass filter ≥ 650 nm.
Notes:The optimal dye concentration and incubation time may vary depending on cell type and application.