Description
Application
- For evaluation of drugs and screening of potential inhibitors to pepsin 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 pepsin activity
Species
- All
Procedures
- 45 min
Size
- 200 tests in 96-well plate
Detection Limit
- NA
Shelf Life
- 6 months
More Details
PEPSIN (EC 3.4.23.1) is a digestive, serine protease that hydrolyzes dietary proteins in many eukaryotic and prokaryotic organisms. Pepsin predominantly cleaves peptide chains at the amino side of the aromatic phenylalanine, tryptophan, and tyrosine amino acids. BioAssay Systems’ QuantiFluo™ Pepsin Inhibitor Assay Kit uses a fluorescein isothiocyanate (FITC)-labeled synthetic substrate. The fluorescein label is highly quenched. Upon digestion by pepsin 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.
