QuantiChrom™ β-Glucuronidase Assay Kit

Protocol SDS

Application

• For quantitative determination of β-glucuronidase enzyme activity in biological samples, e.g. blood, tissue and bacterial samples.

Key Features

• Highly Sensitive. The fluorimetric nature of the kit detects 1×10^-4 U/L to 8 U/L BG activity in 96 well plate with only 10 µL sample within 30 min.
• Versatile. This assay has been validated to work with both animal and E. coli derived β-glucuronidase.
• High-throughput. Homogeneous “mix-incubate-measure” type assay. Can be readily automated to assay thousands of samples per day.

Method

• FL365/450nm

Samples

• Blood, tissue and bacterial samples

Species

• All

Procedures

• 30 min

Size

• 100 tests in 96-well plate

Detection Limit

• 10^-4 U/L

Shelf Life

• 6 months

More Details

BETA-GLUCURONIDASE (BG) is an enzyme which catalyzes the hydrolysis of β-glucuronide bonds. In humans, β-glucuronidase catalyzes the hydrolysis of β-D-glucuronic acids from glycoproteins and mucopolysaccharides such as heparan sulfate. β-glucuronidase activity is important for drug metabolism as drugs are often conjugated to glucuronic acid to make for a more water soluble delivery system, relying on subsequent β-glucuronidase activity to release the active drug. Bilirubin clearance is also facilitated by glucuronidation. GUSB is often used as a reporter gene to track gene expression through β-glucuronidase activity. BioAssay Systems’ fluorimetric BG assay uses a β-D-glucuronide substrate which fluoresces upon hydrolysis with β-glucuronidase and is read at 365/450 nm.

Will this kit detect β-glucuronidase from my organism?

This kit was optimized for mammalian BG, but also works with E.coli BG. The substrate is at pH 5.0 and should be suitable for enzymes with optimal pH in the range of 3-8. We recommend to run an intial test with 30 minute incubation and if results show low activity, extend the incubation to 60 or 90 minutes.

Can I run this assay in a black 384 well plate?

Yes, to use 384 well plate format simply divide all volumes by two, e.g. 5 µL sample or standard, 20 µL substrate, 7.5 µL stop reagent. Otherwise the protocol is the same.

Is sample background fluorescence an issue?

In our testing we did not see noticeable background from tissue lysates or serum samples. Generally samples do not fluoresce at the kit’s wavelength, or are diluted enough to not be an issue. In the rare case that samples do have significant background fluorescence we advise to include a sample blank that uses deionized water in lieu of substrate.

For more detailed product information and questions, please feel free to email or call us at 1-510-782-9988 x 2. For more general information regarding our assays, please refer to Technical Support.

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If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.

– Fast turnaround
– Quality data
– Low cost

Please email or call 1-510-782-9988 x 2 to discuss your projects.