BODIPY-FL 

BODIPY-FL is a green fluorescent dye that can be used to label probes or primers. BODIPY-FL fluorescence can be quenched after interacting with uniquely positioned guanine, making it useful for quantifying specific DNA or RNA molecules. BODIPY-FL-labeled sphingolipid analogs can be used to investigate sphingolipid internalization, transport, and endocytosis in mouse embryonic stem cells. BODIPY-FL-labeled monoterpenes can quickly penetrate bacteria, mammalian, and fungal cells, allowing for the detection of characteristics of a broad spectrum of Gram-positive and Gram-negative bacteria, as well as pathogenic fungi. The maximum absorption wavelength of BODIPY-FL is 505 nm, and the maximum emission wavelength is 513 nm^[1][2][3][4].

Preparation of BODIPY FL-Phospholipid Staining Solution^[3]
1.1 Preparing the Storage Solution
Prepare a 1 mM stock solution of BODIPY FL-ceramide using chloroform/ethanol solvent (19:1 (v/v)).
Note: 1) Transfer the BODIPY FL-Phospholipid storage solution to an amber glass round sample vial and store it in the dark at -20 ℃.
1.2 Preparing the work solution
1) Place the nozzle of the nitrogen evaporator into the amber glass round bottle and spray nitrogen onto the storage liquid until the liquid phase is completely dry; then add 200 µL of anhydrous ethanol to reconstitute the dried BODIPY FL-phospholipid powder.
2) Prepare a 5 µM solution of defatted BSA by dissolving an appropriate amount of defatted BSA in 10 mL of Hanks' buffered salt solution containing 10 mM HEPES (HBSS/HEPES buffer).
3) Add the 200 μL BODIPY FL-sphingolipid ethanol solution prepared in 1) to the 10 mL defatted BSA solution prepared in 2), and mix it on a vortex mixer to get the working solution (5μM BODIPY FL-sphingolipid + 5 μM BSA).
Note: 1) Store the working solution in the dark at -20 ℃.
BODIPY FL - sphingolipid staining
1) Culture mESC cells using ESGRO growth medium, and seed the cells in dishes at a density of 5 × 10⁴ cells/cm², incubating overnight in a 37°C, 5% CO₂ incubator.
2) Remove the growth medium from the overnight cell culture dish and wash the adherent cells three times with HBSS/HEPES buffer.
3) Add the BODIPY FL-sphingolipid working solution to the dish, completely covering the cell growth area, and incubate the cells at 4°C in the dark for 30 minutes.
4) Remove all working liquid and wash the cells with ESGRO growth medium three times.
5) Add RT ESGRO growth medium to cover the cells, and incubate them in the dark at 37°C with 5% CO₂ for 30 minutes.
6) Wash the cells with ESGRO growth medium, and examine and image the stained cells using the LSM 780 ultraviolet confocal microscope.
7) Data analysis (confocal microscopy): Organelles or areas with high levels of BODIPY FL-sphingolipid analogs emit red fluorescence (λem ≥ 590 nm), while organelles or areas with low levels of BODIPY FL-sphingolipid analogs emit green fluorescence (λem = 520-590 nm).

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

292.09

C₁₄H₁₅BF₂N₂O₂

165599-63-3

Solid

Brown to red

516

495

O=C(CCC1=CC=C2C=C3C(C)=CC(C)=[N]3[B+3]([F-])([N-]21)[F-])O

Room temperature in continental US; may vary elsewhere.

4°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]. Leung K. BODIPY-FL-neutravidin-biotin-Cetuximab. 2008 Jan 15 [updated 2008 Feb 25]. In: Molecular Imaging and Contrast Agent Database (MICAD) [Internet]. Bethesda (MD): National Center for Biotechnology Information (US); 2004–2013. PMID: 20641313.  [Content Brief]

  [2]. Kurata S, et al. Fluorescent quenching-based quantitative detection of specific DNA/RNA using a BODIPY((R)) FL-labeled probe or primer. Nucleic Acids Res. 2001 Mar 15;29(6):E34.  [Content Brief]

  [3]. Fan W, et al. Using BODIPY FL-Sphingolipid Analogs to Study Sphingolipid Metabolism in Mouse Embryonic Stem Cells. Bio Protoc. 2022 Nov 20;12(22):e4555.  [Content Brief]

  [4]. Guseva GB, et al. Design, Spectral Characteristics, and Possibilities for Practical Application of BODIPY FL-Labeled Monoterpenoid. ACS Appl Bio Mater. 2021 Aug 16;4(8):6227-6235.  [Content Brief]