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].

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.

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

370.63

C₁₆H₂₂BClF₂N₄O

209541-26-4

Solid

Orange to red

510

502

[F-][B+3]1([N-]2C(C=C(C(C)=C3)[N]1=C3C)=CC=C2CCC(NCCN)=O)[F-].Cl

Room temperature in continental US; may vary elsewhere.

-20°C, sealed storage, away from moisture and light

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

• [1]. Cornelius M, et al. Detection and separation of nucleoside-5'-monophosphates of DNA by conjugation with the fluorescent dye BODIPY and capillary electrophoresis with laser-induced fluorescence detection. Electrophoresis. 2005 Jun;26(13):2591-8.  [Content Brief]

  [2]. Zinellu A, et al. Capillary electrophoresis with laser-induced fluorescence detection for ATP quantification in spermatozoa and oocytes. Anal Bioanal Chem. 2010 Nov;398(5):2109-16.  [Content Brief]

  [3]. Cornelius MG, et al. RNA analysis by MEKC with LIF detection. Electrophoresis. 2007 Nov;28(21):3901-7.  [Content Brief]