Rhodamine B labeled Polymyxin B Ready Made Solution
SIGMA/SBR00036 - For fluorescent microbial imaging, 1.1 mg/mL in water
Synonym: SRB-PMB
Product Type: Chemical
Escherichia coli cells stained with Rhodamine B labeled Polymyxin B E. coli cells were harvested at mid-log phase. Pellets were washed twice with 1X PBS and incubated at a final concentration of 1.1 μg/mL Rhodamine B labeled Polymyxin B solution at room temperature for ~2 min, followed by 4 washes with PBSX1. Pellets were finally reconstituted to 100 μL with 1X PBS. Images were acquired immediately after staining using Olympus BX53F2 upright microscope (X40X2 magnification). Image depicts Rhodamine labeled Polymyxin B staining, excitation –540 to 550 nm, emission 575 to 625 nm (CY3 filter).
Escherichia coli cells stained with Rhodamine B labeled Polymyxin B and DAPI (Cat. No. MBD0015) E. coli cells were harvested at mid-log phase. Pellets were washed twice with 1X PBS and incubated at a final concentration of 1.1 μg/mL Rhodamine B labeled Polymyxin B solution at room temperature for ~2 min, followed by 4 washes with PBSX1. Pellets were finally reconstituted to 100 μL with 1X PBS. DAPI (Cat. No. MBD0015) was added to a final concentration of 1 μg/m. Images were acquired immediately after staining using Olympus BX53F2 upright microscope (X40X2 magnification). Image depicts Rhodamine labeled Polymyxin B staining, excitation –540 to 550 nm, emission 575 to 625 nm (CY3 filter). DAPI was detected with excitation –360 to 370 nm, emission 420 to 460 nm (DAPI filter)
| concentration | 1.1 mg/mL in water |
| Quality Level | 100  |
| storage temp. | −20°C |
| Analysis Note: | • Rhodamine B labeled polymyxin B Ready Made Solution is light sensitive. • It is recommended to avoid freeze-thaw cycles of Rhodamine B labeled polymyxin B Ready-Made Solution • Rhodamine B labeled polymyxin B Ready Made Solution (1.1mg/mL) can be diluted 1:100-200 in PBSX1 (Product# D8537) to achieve 0.55-1.1μg/mL final concentration for staining. The above concentration of Rhodamine B labeled polymyxin B was used for Escherichia coli staining (see image) • Fluorescence Microscopy application: Rhodamine B labeled polymyxin B Ready Made Solution excitation (Ex) wavelength is 550-560nm resulting in emission (Em) range of 580-610nm (λmax=590-600nm) |
| 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. Additional possible applications for Rhodamine B labeled Polymyxin B:• Evaluate LPS binding by FRET (fluorescence resonance energy transfer) studies to form a specific complex between fluorescein (FITC)-tagged LPS and Polymyxin B, indicating direct binding of the antibiotic to the LPS • Can be applied as a fluorescent probe to study polymyxin mode of action and its pharmacokinetics • to assess the mitochondrial function of polymyxin B in a kidney cell line (LLC-PK1), suggesting that it changes the mitochondrial membrane polarization. |
| General description: | Rhodamine B labeled polymyxin B (SRB-PMB) is a fluorescent derivative of Polymyxin B. Rhodamine B polymyxin B is an additional fluorescent derivative of Polymyxin B, along with Dansyl labeled polymyxin B (Product # SBR00029). Both products′ mode of action is in accordance with Polymyxin B activity. Fluorescent antibiotics are obtained by synthetic conjugation of an antibiotic to a fluorophore. |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |