EnzyFluo™ Uricase Assay Kit

Protocol SDS

Application

• Uricase activity in biological samples such as cell culture, non-human biofluids, cell extracts, and plant tissue extracts.

Key Features

• Sensitive. Use 10 μL samples. Linear detection range: fluorimetric assay 0.3 – 100 U/L Uricase.
• Fast. Run time is 10 minutes for rapid results
• Convenient. Room temperature “mix-and-read” procedure can be readily automated for high-throughput assay of thousands of samples per day.

Method

• FL530/585nm

Samples

• Biological samples

Species

• All (except hominids)

Procedures

• 10 min

Size

• 100 tests in 96-well plate

Detection Limit

• 0.3 U/L

Shelf Life

• 6 months

More Details

URICASE or URATE OXIDASE (E.C. 1.7.3.3) initiates the oxidation of uric acid, eventually yielding allantoin. Uricase activity is found throughout plants, animals, and bacteria, but it is not found in hominids. As a result, humans develop gout when the levels of uric acid become too high. BioAssay Systems’ method provides a simple and high-throughput assay for measuring uricase enzyme activity. In this assay, uric acid is enzymatically converted to allantoin, releasing H₂O₂. The resulting H₂O₂ reacts with a specific dye to form a pink-colored product. The change in fluorescence intensity at 530/585 nm is directly proportional to the uricase activity in the sample.

How do I prepare plant extracts for uricase analysis?

We used a Dounce tissue homogenizer with ice-cold 100 mM Tris-HCl (pH 7.5) and ground up etiolated pea shoots and roots then assayed the extracts fresh. Protease inhibitors can be added, as well.

How do I prepare animal tissue extracts for uricase analysis?

We recommend using exsanguinated tissues then homogenizing in 50 mM Tris-HCl (pH 7.5), 150 mM KCl, 2 mM EDTA with a Dounce homogenizer.

Can I use Optical Density to measure uricase activity with this kit?

Yes, the dye in the kit does have an Optical Density peak at 570nm, but we found that the activity was more linear at lower concentrations of uricase while employing fluorescence. If you choose to use OD, you may need to use a non-linear fit for the standard curve.

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.