QuantiFluo™ Chymotrypsin Inhibitor Assay Kit

Protocol SDS

Application

• For evaluation of drugs and screening of potential inhibitors to chymotrypsin 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 >0.8.

Method

• FL485/530 nm

Samples

• Compounds that affect chymotrypsin protease activity.

Species

• All

Procedures

• 45 min

Size

• 100 tests in 96-well plate.

Detection Limit

• NA

Shelf Life

• 6 months

More Details

CHYMOTRYPSIN (EC 3.4.21.1) is a digestive, serine protease that hydrolyzes dietary proteins in many eukaryotic and prokaryotic organisms. Chymotrypsin predominantly cleaves peptide chains at the carboxyl side of the aromatic phenylalanine, tryptophan, and tyrosine amino acids. BioAssay System’s QuantiFluo™ Chymotrypsin Inhibitor Assay Kit uses a fluorescein isothiocyanate (FITC)-labeled synthetic substrate. The fluorescein label is highly quenched. Upon digestion by chymotrypsin 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 λ_ex/em = 485/530 nm. Inhibition is determined by the decrease in fluorescence.

Is chymotrypsin enzyme or inhibitor included in the kit?

The customer must provide their own enzyme and inhibitor. We used purified chymotrypsin (MedChemExpress, cat# HY108910) and trypsin soybean inhibitor (Cayman Chemical, cat# 14502)

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 chymotrypsin solution need to be prepared fresh for each assay run? I know that, after some time, proteases like chymotrypsin may begin to digest themselves.

To prevent autodigestion, pure chymotrypsin solutions can be stored long term at -80°C under acidic conditions (1 mM HCl, pH 3.0). 2 mM CaCl₂ can be added to stabilize the stock solution.

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.