BODIPY 581/591 C11,IC-0153066

BODIPY 581/591 C11 is a BODIPY borofluoroprene derivative with good light stability and low fluorescence artifacts. BODIPY 581/591 C11 can be used for study lipid peroxidation and antioxidant properties in living cells, or detect ferroptosis by reaction with hydroxyl radicals. BODIPY 581/591 C11 is emitted at 591 nm (reduced prototype), or redshifted to 510 nm (oxidized type). The excitation wavelengths were 581 nm (reduced prototype) and 500 nm (oxidized type)^[1].

Usage

I. Stock Solution Preparation (10 mM)

1. Dissolution Method  

1.1 Weigh 1 mg BODIPY 581/591 C11 and dissolve it in 0.1983 mL DMSO with thorough mixing.  

1.2 Calculation Basis: Assuming the molecular weight is M (refer to the reagent manual), 1 mg / M = 10 mmol/L ¡Á 0.0001983 L ¡ú Verify M ¡Ö 504.5 g/mol (adjust the volume if the actual molecular weight differs).  

2. Storage Conditions  

Aliquot into small volumes and store protected from light at -20¡ãC or -80¡ãC. Avoid repeated freeze-thaw cycles (single-use aliquots are recommended).  

II. Working Solution Preparation (2¨C10 ¦ÌM)

1. Dilution Method  

1.1 Mix X ¦ÌL of 10 mM stock solution + Y ¦ÌL serum-free medium/PBS.  

1.2 Examples:  

1.2.1 To prepare 10 ¦ÌM working solution: 1 ¦ÌL stock + 999 ¦ÌL medium (1:1000 dilution).  

1.2.2 To prepare 2 ¦ÌM working solution: 1 ¦ÌL stock + 4999 ¦ÌL medium (1:5000 dilution).  

2. Notes  

2.1 Prepare freshly and protect from light.  

2.2 Optimize the concentration based on cell type and experimental needs (e.g., toxicity testing).  

III. Cell Staining Protocol

1. Cell Preparation

1.1 Suspension Cells: Centrifuge (1000g, 5 min) ¡ú Wash twice with PBS.  

1.2 Adherent Cells: Detach with trypsin ¡ú Centrifuge ¡ú Wash twice with PBS.  

2. Staining and Detection

1. Resuspend cells in 1 mL working solution (2¨C10 ¦ÌM) and incubate at room temperature, protected from light, for 30 min.  

2. Centrifuge (400g, 4¡ãC, 3¨C4 min) ¡ú Discard supernatant ¡ú Wash twice with PBS.  

3. Resuspend in serum-free medium/PBS and proceed immediately with:  

Fluorescence microscopy or flow cytometry analysis (excitation/emission reference: ~581/591 nm; instrument calibration required).  

IV. Key Considerations

1. Storage and Stability  

   - Avoid repeated freeze-thaw cycles of the stock solution (¡Ý3 cycles may cause degradation).  

   - Working solutions should ideally be used on the same day, as prolonged light exposure may cause photodegradation.  

2. Experimental Optimization  

   - Concentration gradient test: Compare 2 ¦ÌM, 5 ¦ÌM, and 10 ¦ÌM to avoid over-staining or high background.  

   - Incubation time: Test 20¨C60 min to minimize nonspecific binding.  

3. Safety Precautions  

   - Wear a lab coat and gloves; handle DMSO in a fume hood.  

   - Dispose of waste according to organic solvent regulations.  

4. Control Setup  

   - Negative control: Unstained cells.  

   - Solvent control: Cells treated with an equivalent volume of DMSO (to rule out solvent effects).  

V. Frequently Asked Questions

- Q1: Weak staining signal?  

A1 :Increase working solution concentration (e.g., 10 ¦ÌM) or extend incubation time (up to 45 min).  

- Q2: High background?  

A2: Increase PBS washes (3¨C4 times) or reduce working solution concentration.  

Rerference

[1]. Gregor P C Drummen, et al. C11-BODIPY(581/591), an oxidation-sensitive fluorescent lipid peroxidation probe: (micro)spectroscopic characterization and validation of methodology. Free Radic Biol Med. 2002 Aug 15;33(4):473-90.

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