Hexosaminidase activity

Hexosaminidase activity

The β-N-Acetylglucosaminidase Activity Fluorometric Assay Kit enables highly sensitive and specific quantification of hexosaminidase activity in a wide range of biological samples, including tissues, cells, serum, and urine. Hexosaminidase, particularly β-N-Acetylglucosaminidase (β-NAG), is a critical lysosomal enzyme found in organs such as the kidneys, liver, lungs, and in immune cells like mast cells. Abnormal activity levels of this enzyme have been linked to several pathological conditions, including lysosomal storage disorders like Tay-Sachs and Sandhoff disease, as well as chronic inflammation and immune dysfunctions. By providing a reliable method to measure enzyme activity, this kit supports research into disease mechanisms, diagnosis, and therapeutic monitoring.

The assay operates on a straightforward yet powerful principle: it utilizes the non-fluorescent substrate 4-Methylumbelliferyl N-acetyl-β-D-glucosaminide (4-MUAG), which is cleaved by β-NAG to release the fluorescent product 4-Methylumbelliferone (4-MU). The fluorescence can be quantitatively measured using an excitation wavelength of 365 nm and an emission wavelength of 445 nm. This allows for real-time kinetic monitoring of enzymatic activity with high accuracy and reproducibility. The linear detection range and optimized reaction conditions (typically 30 minutes at 37°C using 20 µL samples) ensure that both low- and high-abundance samples yield reliable data.

Designed for flexibility and efficiency, the kit is compatible with high-throughput screening (HTS) platforms, allowing researchers to process thousands of samples per day. Its ready-to-use components—including a substrate, standard, positive control, stop reagent, and assay buffer—are conveniently packaged for either 100 or 200 reactions. The kit is shipped on ice and maintains a 12-month shelf life when stored at -20°C. To ensure consistent results, it is recommended that substrate addition and reaction termination are performed rapidly and uniformly, preferably using a multi-channel pipettor. This assay is an indispensable tool for researchers studying lysosomal function, metabolic disorders, and immune-related conditions.