BODIPY FL-EDA

BODIPY FL-EDA is a widely used fluorescent dye for quantitative analysis of nucleotides. BODIPY FL-EDA is an aliphatic amine analog that can react with aldehydes and ketones. BODIPY FL-EDA can be used to detect both modified and unmodified deoxynucleotides and to determine DNA damage and genomic DNA methylation through capillary electrophoresis with laser-induced fluorescence (CE-LIF). Additionally, it can be used for quantifying intracellular ATP levels. The excitation wavelength is 500 nm, and the emission wavelength is 510 nm^[1][2][3]. In Vitro: 1.1 Preparation of Stock Solution
Prepare a 22 mmol/L BODIPY FL-EDA solution in DMSO (HY-Y0320) by dissolving 8.15 mg of BODIPY FL-EDA in 1 mL of DMSO.
Note: It is recommended to aliquot the BODIPY FL-EDA stock solution and store it protected from light at -20°C or -80°C.
1.2 Quantitative Detection of ATP in Sperm and Oocytes Using BODIPY FL-EDA Labeling
1.2.1 Sample Derivatization: Mix 10 μL of lysed sperm or oocyte samples with 40 μL of 1.4 mol/L EDC (dissolved in 50 mmol/L HEPES (HY-D0857), pH 7.5) and 5 μL of the 22 mmol/L BODIPY FL-EDA stock solution. Incubate the mixture in the dark at 25°C for 24 hours.
1.2.2 Perform quantitative detection of ATP using capillary electrophoresis with laser-induced fluorescence detection (CE-LIF)
(1) CE Separation: Dilute the derivatized sample 40 times with water before injecting it into the capillary electrophoresis instrument. Under the influence of a high-voltage electric field, BODIPY-ATP will migrate in the capillary based on its charge-to-mass ratio.
(2) LIF Detection: At the capillary outlet, use a laser to excite the fluorescence of BODIPY-ATP. The excitation wavelength is 500 nm, and the emission wavelength is 510 nm. The emitted fluorescence is detected by a photomultiplier tube and converted into an electrical signal.
(3) Standard Curve Preparation: Perform CE-LIF analysis using a series of ATP standards with known concentrations to generate a standard curve correlating fluorescence intensity with ATP concentration.
(4) Data Analysis: Calculate the ATP concentration in the samples based on the fluorescence intensity using the standard curve.