BODIPY FL-EDA

Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
1.1 Preparation of Stock Solution
Prepare a 22 mmol/L BODIPY FL-EDA solution in 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.
2 Quantitative Detection of ATP in Sperm and Oocytes Using BODIPY FL-EDA Labeling
2.1 Sample Derivatization: Mix 10 μL of lysed sperm or oocyte samples with 40 μL of 1.4 M EDC (dissolved in HEPES (HY-D0857), pH 7.5) and 5 μL of the 22 mM BODIPY FL-EDA stock solution. Incubate the mixture in the dark at 25°C for 24 hours.
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.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.