QuantiFluo™ Trypsin Inhibitor Assay Kit
- For evaluation of drugs and screening of potential inhibitors to trypsin proteases.
- 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.
- FL485/530nm or FP485/530nm
- Compounds that affect trypsin activity
- 45 min
- 100 tests in 96-well plate
- 6 months
More DetailsTRYPSIN (EC 184.108.40.206) 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.
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