Through fine-tuning of molecular structure, a new generation of fluorescent substrates is breaking through traditional performance bottlenecks.

DiFMUP (6,8-Difluoro-4-methylumbelliferyl phosphate) is a representative of this trend—significantly overcoming the shortcomings of traditional 4MU, such as low luminescence efficiency in acidic/neutral environments and susceptibility to photobleaching, making it an ideal choice for a new generation of high-reliability detection.

Its core advantage stems from the structural optimization of its hydrolysis product, DiFMU: The introduction of fluorine atoms at the 6,8- positions significantly reduces the pKa of the phenolic hydroxyl group (~4.7 vs. ~7.9 for 4MU), making it easier to deprotonate within the pH 6~8 range. This change brings comprehensive product improvements:

This is not just a performance upgrade, but a comprehensive improvement in detection reliability.

DiFMUP serves as a fluorescent phosphatase substrate. When it encounters a specific phosphatase, the phosphate ester bond is hydrolyzed, releasing the fluorescent compound 6,8-difluoro-4-methylumbelliferone. By detecting the fluorescence intensity of this substance, the activity level of the phosphatase can be quantified, thereby providing a basis for related enzymology research and diagnostics.

1. Enzyme Activity Detection: Can be used to detect the activity of various phosphatases, including acid phosphatase, alkaline phosphatase, and protein tyrosine phosphatases.

2. High-Throughput Screening: Its high sensitivity and suitability for continuous detection make it an ideal tool for screening phosphatase inhibitors or activators in drug discovery.

3. Immunoassay Analysis: As a fluorescent substrate for alkaline phosphatase-labeled antibodies, it is widely used in immunological detection methods such as ELISA and Western Blot, enabling high-sensitivity protein detection.

✅ Ultimate Sensitivity: Higher quantum yield and optimal ionization state deliver signal intensity several times that of 4MU, easily capturing weak signals.

✅Stable Signal: Extremely low pKa value completely eliminates interference from pH fluctuations, ensuring highly reproducible data and smoother kinetic curves.

✅ pH-Worry-Free: Stable fluorescence output within pH 6.0~8.0 range, no need for strict buffer control, ensuring result repeatability and accuracy.

✅ Perfect Compatibility: Spectral wavelengths remain almost unchanged, no need to adjust detection parameters, zero-cost upgrade, immediately enjoy comprehensive performance improvement.