Analysis Note

Aliquots of the DMSO solution can be stored at -20°C, protected from light for at least 1 month. Fluorescent microscopy application: Vancomycin -FITC has excitation/emission wavelength at 495 nm/515 nm. It is recommended to dissolve Vancomycin-FITC in DMSO (Dimethyl Sulfoxide) to a concentration of 1mg/mL. Recommended working concentration for Bacillus cereus in fluorescent microscopy imaging application is 25µg/mL.

Application

Fluorescent antibiotics can be used for many applications including: Antimicrobial resistance research. Bacterial visualization and imaging. Parent antibiotic mode of action research and new antibiotic discovery. Toxicity studies. Research of bacterial infections and tracking its uptake in vivo.

Biochem/physiol Actions

Vancomycin inhibits cell wall synthesis at the level of peptidoglycan biosynthesis. Vancomycin binds with high affinity and specificity to the terminal D-Ala-D-Ala of the growing peptide chain during cell wall synthesis. The binding inhibits transpeptidase activity, which prevents further elongation and cross-linking of the peptidoglycan matrix. Vancomycin-FITC has fluorescein attached to the amine group on the disaccharide moiety of the vancomycin molecule. This disaccharide moiety does not play any role in D-Ala-D-Ala binding site, affording a comparable binding affinity of Vancomycin-FITC to Gram-positive bacteria as the parent drug. However, a minimum inhibitory concentration (MIC) decrease of about two orders of magnitude in comparison to Vancomycin was observed. This activity decrease can be linked to repulsive interactions between the negatively charged Fluorescein moiety and the anionic environment of the peptidoglycan.

General description

FITC labeled Vancomycin (Vancomycin-FITC), is a Fluorescein conjugated Vancomycin, with a similar mode of action as Vancomycin. Vancomycin -FITC can be used to detect Gram-positive bacteria by common fluorescent instruments. Vancomycin is a glycopeptide antibiotic, used to treat gram-positive infections. Fluorescent antibiotics are obtained by synthetic conjugation of an antibiotic to a fluorophore.