PFB-FDG (solution)

Guidelines (The following is our recommended experimental protocol. This protocol serves only as a reference guide, and specific operations should be adjusted according to your actual needs).
β-Galactosidase Activity Assay^[6]:
1. Incubate 2 × 10⁵ cells in serum-free DMEM medium containing 25 mM Hepes (pH 7.4) supplemented with 50 μg/mL PFB-FDG at 37°C for 2 h. (Taking monocytes as an example, cell concentration can be adjusted for different cell types)
2. Wash the cells once with pre-chilled PBS.
3. Analyze the cells using a flow cytometer (excitation wavelength Ex=485 nm, emission wavelength Em=535 nm).

PFB-FDG (solution) (20-40 μM; pre-incubated with conduritol B epoxide for 30 min) detects significant lysosomal GBA deficiency (residual activity is 2-4% of normal levels) in fibroblasts from LIMP2-deficient AMRF patients, while significant residual GBA activity is detected in lymphocytes (35% of normal levels), monocytes (47% of normal levels), and monocyte-derived macrophages (32% of normal levels) from AMRF patients^[5].
PFB-FDG (solution) (3 h) is a non-leaky fluorescent substrate that can be used to detect functional β-galactosidase synthesized inside POPC/cholesterol/DSPE-PEG₅₀₀₀ liposomes. Enzyme activity is determined by measuring green fluorescence via flow cytometry (FACS) after incubation at 37°C for 3 h^[1].
PFB-FDG (solution) enables the detection of senescence-associated β-galactosidase activity via hydrolysis. Results show that senescence-associated β-galactosidase activity is significantly higher in freshly isolated human CD34⁺ CD133⁺ hematopoietic progenitor cells from middle-aged donors than in those from young donors^[2].
PFB-FDG (solution) (2 h) can be activated by endogenous SA-β-gal in viable human U251.JMJD3wt glioma cells, generating detectable green fluorescence after incubation at a concentration of 3 mg/mL for 2 h^[3].
PFB-FDG (solution) (50 μM; overnight) is a viable fluorescent substrate for the β-galactosidase reaction inside microfluidics-derived GUVs. It undergoes efficient hydrolysis during overnight incubation at 37 °C, while the size of GUVs remains stable^[4].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.