Beta-Glucuronidase is a hydrolytic enzyme that breaks down carbohydrates by cleaving the terminal beta-D-glucuronic acid residue from the non-reducing terminus of a mucopolysaccharide chain. In humans, these enzymes are found in the lysosome of many tissues. Loss of beta-Glucuronidase activity results in a metabolic disease known as Sly syndrome. As the expression and activity of beta-Glucuronidase can vary substantially between tissue types and disease states, these enzymes have been used to achieve targeted activation of glucuronidated prodrugs.

Beta-Glucuronidase Activity Assay Kit provides a simple, reproducible, and sensitive tool of measuring the activity of beta-Glucuronidase in cell lysates, plasma, serum, urine, saliva and other biological liquid samples. In this assay, beta-Glucuronidase cleaves a synthetic specific substrate releasing a fluorophore. The fluorescence signal is measured at Ex/Em = 360/445 nm. The increase in fluorescence intensity is directly proportional to beta-Glucuronidase activity in the sample.

• Microplate reader capable of measuring fluorescence intensity at Ex/Em = 360/445 nm
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Incubator at 37°C
• Tubes to prepare sample dilutions
• 15 mL and 50 mL conical tubes
• MilliQ water or another type of double distilled water (ddH₂O)

1. Add 40 µL of samples, positive control, substrate background to the 96-well microtiter plate (black bottom).
2. Add 60 µL of standard to the 96-well microtiter plate.
3. Add 20 µL diluted β-Glucuronidase substrate solution to each well, but not the standard wells. Carefully shake the plate for a few seconds to mix.
4. Incubate at 37°C for 30 minutes with mixing, protect from light.
5. Add 200 µL Stop Solution. Put the plate on a shaker for 2 minutes to mix well (protect from light).
6. Read the fluorescence signal with a fluorescence plate reader at Ex/Em = 360/445 nm.