Rhodamine B labeled Ramoplanin
SIGMA/SBR00047 - For fluorescent microbial imaging
Empirical Formula (Hill Notation): C146H184ClN23O46S2
Molecular Weight: 3096.73
Linear Formula: C146H184ClN23O46S2
Product Type: Chemical
Bacillus subtilis cells staining with Rhodamine B labeled Ramoplanin.
B. subtilis cells were harvested at mid-log phase. Pellets were washed with 1X PBS and incubated at a final concentration of 5065 μM Rhodamine B labeled Ramoplanin solution. The pellets were washed three times with 1X PBS. Pellets were finally reconstituted to 100 μL with 1X PBS.
Images were acquired immediately after staining using Olympus BX53F2 upright microscope (40X2 magnification). Image depicts Rhodamine labeled Ramoplanin staining, excitation/emission wavelength at 565 nm/585 nm.
B. subtilis cells were harvested at mid-log phase. Pellets were washed with 1X PBS and incubated at a final concentration of 5065 μM Rhodamine B labeled Ramoplanin solution. The pellets were washed three times with 1X PBS. Pellets were finally reconstituted to 100 μL with 1X PBS.
Images were acquired immediately after staining using Olympus BX53F2 upright microscope (40X2 magnification). Image depicts Rhodamine labeled Ramoplanin staining, excitation/emission wavelength at 565 nm/585 nm.
| Quality Level | 100  |
| specific activity | Antibiotic (bacterial staining) |
| storage temp. | −20°C |
| Analysis Note: | • Fluorescent microscopy application: Rhodamine B labeled Ramoplanin has excitation/emission wavelength range at 550-560 nm/590-600 nm. • It is recommended to dissolve the Rhodamine B labeled Ramoplanin in DMSO to a concentration of 2 mg/mL. • The recommended working concentration in fluorescent microscopy imaging application is 10μM. The mentioned concentration was used for Bacillus subtills staining (see image). • Aliquots of the DMSO solution can be stored at −20 °C, protected from light for at least one month. |
| 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 • Diagnosis of bacterial infections and tracking their uptake in vivo |
| Biochem/physiol Actions: | Ramoplanin blocks the bacterial cell wall biosynthesis by specifically binding to the first sugar of lipid II, N-Acetylmuramic acid, thus interfering with the peptidoglycan production. Its mode of action is different from the d-Ala-d-Ala that are targeted by vancomycin and shows no cross-resistance with other glycopeptides. Ramoplanin is a potent antibiotic with an MIC value of =1 μg /ml and broad-spectrum activity against Gram-positive pathogens both in vitro and in vivo, including Enterococci, Staphylococci, Bacilli, Streptococci, Listeria monocytogenes, and Gram-positive anaerobes such as Clostridrium difficile. However, Rhodamine B labeled Ramoplanin, presents an MIC decrease in the magnitude of two to five orders in bacterial activity in comparison to the parent Ramoplanin. |
| General description: | Rhodamine B labeled Ramoplanin is a fluorescent derivative of Ramoplanin, with a similar mode of action as Ramoplanin. It allows fluorescent labeling of Gram-positive bacteria and their detection by common fluorescent instruments. Ramoplanin is a lipoglycodepsipeptide antibiotic that is produced by the fermentation of Actinoplanes species. Fluorescent antibiotics are obtained by a synthetic conjugation of an antibiotic to a fluorophore. |
| activity | specific activity: Antibiotic (bacterial staining) |
| Storage Temp. | −20°C |