QuantiFluo™ Trypsin Inhibitor Assay Kit

Protocol SDS

Application

• For evaluation of drugs and screening of potential inhibitors to trypsin proteases.

Key Features

• Safe. Non-radioactive assay.
• Homogeneous and convenient. Homogenous “Mix-incubate-measure” type assay. No wash and reagent transfer steps are involved.
• Robust and High-throughput. Can be readily automated to assay thousands of samples per day. Robust assay with a Z’ factor of > 0.7.

Method

• FL485/530nm or FP485/530nm

Samples

• Compounds that affect trypsin activity

Species

• All

Procedures

• 45 min

Size

• 100 tests in 96-well plate

Detection Limit

• NA

Shelf Life

• 6 months

More Details

TRYPSIN (EC 3.4.21.4) is a digestive, serine protease that hydrolyzes dietary proteins in many eukaryotic and prokaryotic organisms. Trypsin predominantly cleaves peptide chains at the carboxyl side of lysine and arginine amino acids, but not before proline. BioAssay Systems’ QuantiFluo™ Trypsin Inhibitor Assay Kit uses a fluorescein isothiocyanate (FITC)-labeled synthetic substrate. The fluorescein label is highly quenched. Upon digestion by trypsin present in the sample, the substrate is cleaved into smaller peptides, which abolishes the quenching of the fluorescence label. The fluorescence or fluorescence polarization (FP) of the FITC-labeled fragments is measured at λex/em = 485/530 nm. Inhibition is determined by the decrease in fluorescence.

Is trypsin enzyme or inhibitor included in the kit?

The customer must provide their own enzyme and inhibitor. We used purified trypsin (Sigma Aldrich, cat# T8003) and trypsin inhibitor (Phenylmethanesulfonyl fluoride (PMSF), Sigma Aldrich Cat# P7626).

Can the substrate be refrozen after use?

While the substrate can handle a few freeze-thaw cycles, if the customer plans to run many tests, it would be best to aliquot the substrate into light-protected tubes and thaw as needed.

Does trypsin solution need to be prepared fresh for each assay run? I know that, after some time, trypsin begins to digest itself.

Trypsin can be stored long term in solution at -80°C under acidic conditions (1 mM HCl, pH 3.0). This should reduce the speed of self-digestion significantly.

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.

No citations for this new product. Please check back later. You may click here to check if citations are available, but are not listed here yet.

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.