Filipin complex

Filipin complex is a potent polyene macrolide antifungal antibiotic. Filipin complex inserts into membranes and sequester cholesterol into complexes and inhibits PRRSV entry. The Filipin complex consists of about 75.8% Filipin III (HY-N6718), 10.8% Filipin IV, 9.1% Filipin II, and 1.2% Filipin I (Ex/Em = 380/430 nm)^[1][2][3]. In Vitro:1. Solution preparation^[4]
1.1 Preparation of storage solution
Solvent: DMSO
Concentration: 50 mg/mL is recommended (e.g. weigh 50 mg Filipin complex and dissolve in 1 mL solvent).
Storage: After aliquoting, store at -20°C or -80°C away from light and avoid repeated freezing and thawing.
1.2 Preparation of working solution
Dilute to 50 μg/mL with PBS or serum-free cell culture medium (optimized according to the experiment).
Note: The working solution should be prepared and used immediately, and protected from light.

For neuronal cholesterol levels staining.
1. The staining sections are washed three times with PBS and incubated with 1.5 mg/mL glycine for 10 min at room temperature.
2. Then sections are stained with 0.05 mg/mL Filipin complex in PBS containing 10% FBS for 2 h and rinsed for three times with PBS.
3. Use an Olympus FV10i confocal microscope for image (Ex/Em = 380/430 nm).

Filipin complex (2 μg/mL; 1+48 h) inhibits PRRSV entry with respiratory syndrome virus (PRRSV) genome copies in infected cells reduced by 61% in MARC-145 cells^[2].