General descriptionBacterial Peptidoglycan is a polymer consisting of glycans and short peptides, a multilayer rigid structure that forms the cell wall. D-sugars and D-amino acids (DAA) are metabolically incorporated into the bacterial cell. It was recently discovered that transpeptidases enzymes catalyze the metabolic incorporation of exogenous D-sugars and DAAs with almost no restriction of the side-chain identity.
Modification of a D-sugar, such as D-Galactosamine, with molecular fluorophores provides fluorescent probes that can efficiently label in situ peptidoglycan in diverse bacterial species.
Rhodamine-labelled D-Galactosamineis a fluorescent derivative of D-Galactosaminethat is obtained by a synthetic conjugation of D-Galactosamineto the fluorophore. Rhodamine-labelled D-Galactosamine has high biocompatibility and suitability for labeling peptidoglycans in live bacteria. Additionally, it can be used in tandem with other stains such as FITC-labelled Alanine (#SBR00049) to distinguish between different bacteria. Other compatible products useful for live staining include: Rhodamine B labelled Polymyxin B, and Ramoplanin, FITC Labelled D-Lysine, Alanine or Vancomycin and Dansyl labelled polymyxin B (catalog numbers: SBR00036, SBR00047, SBR00050, SBR00049, SBR00028, SBR00029).
Application
Fluorescent labeled sugars can be used for many applications including:
- Bacterial cell wall morphology
- Bacterial cell wall formation or remodeling activity
- Bacterial viability/activity
- Identify bacterial activity on surfaces or in substances
- Differentiation between various bacterial strains according to their incorporation profile of different D amino acids and sugars
Analysis Note
- Fluorescent microscopy application: Rhodamine-labelled D-Galactosamine has excitation/emission wavelength range at 542-582/604-644 nm.
- The recommended working concentration in fluorescent microscopy imaging application is between 250-500 µM in working medium.
- Aliquots of the DMSO solution can be stored at -20⁰C, protected from light for at least one month.