| Field | Specification |
|---|---|
| Mfr No | |
| Alternative Names | Bone marrow serine protease, Elastase-2, Human leukocyte elastase (HLE), Medullasin, PMN elastase, ELANE, ELA2 |
| Product Type | |
| Shipping | |
| Storage | |
| UniProt # |
Scientific Background
Elastase is a serine protease from the chymotrypsin family involved in the degradation of elastin and some viral proteins. Elastase exists as pancreatic and neutrophil elastase, which differ in their specificity. Neutrophil elastase is also known as ELA2 or polymorphonuclear leukocyte elastase and is present in azurophilic granules. Upon neutrophil activation, degranulation occurs and serine proteases, such as elastase, are released into the extracellular space to degrade the matrix proteins of pathogens. Alterations in the levels of elastase, or dysfunction of its endogenous inhibitors, result in alpha-1 antitrypsin deficiency (A1AD), emphysema, cyclic neutropenia, abdominal aortic aneurysms (AAA) and chronic obstructive pulmonary disease (COPD), arthritis and psoriasis. The use of neutrophil elastase inhibitors, such as elafin, and the development of new inhibitors with fewer side effects may benefit patients suffering from elastase-related diseases.
Product Description
The Neutrophil Elastase Inhibitor Screening Assay Kit is a fluorogenic assay designed to measure the activity of elastase for screening and profiling applications. The elastase assay kit comes in a convenient 96-well or 384-well format and contains enough purified elastase, fluorochrome-conjugated substrate, and PR-02 buffer for 100 or 400 reactions. This kit also includes Sivelestat as control inhibitor. PR Substrate 3 is a fluorogenic peptide substrate for neutrophil elastase. In the conjugated form the energy emitted from the fluorochrome AMP is quenched. Proteolysis releases AMP and fluorescence is emitted. The increase in fluorescence is directly proportional to elastase activity.
Methodology
Fluorogenic
Applications
Screen small molecule inhibitors in high throughput screening (HTS) applications.
Safety & Handling
For Research Use Only (RUO). Biosafety Level: Not applicable (BSL-1). Note: Troubleshooting Guide: Visit bpsbioscience.com/assay-kits-faq for detailed troubleshooting instructions. For all further questions, please email support@bpsbioscience.com Consult the Safety Data Sheet (SDS) before use.
Avoid freeze/thaw cycles.This assay uses a fluorogenic or chromogenic peptide substrate that is cleaved by neutrophil elastase (NE), releasing a detectable signal proportional to enzyme activity. Inhibitor compounds reduce substrate cleavage, producing a dose-dependent decrease in signal. IC50 values are calculated from percent inhibition curves using non-linear regression.
Yes. This kit is available in both 96-well and 384-well formats, with the 384-well format (82090-2) specifically designed for high-throughput compound library screening. Miniaturized volumes reduce reagent consumption per data point and enable automated liquid handling for large-scale inhibitor screening campaigns.
The kit typically includes positive control (active NE enzyme), substrate, and assay buffer to establish a baseline signal. Researchers should include a known NE inhibitor as a positive inhibition control and a no-enzyme background well as a negative control in each plate to enable data normalization and Z'-factor calculation for HTS assay qualification.
This kit is primarily designed for compound screening using recombinant NE. Measurement of NE activity in complex biological samples (e.g., bronchoalveolar lavage, sputum) may be possible but requires optimization to account for endogenous matrix effects, protease inhibitor carry-over, and background fluorescence. Validate sample compatibility before reporting data from biological matrices.
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- Custom Configurations: Alternative formulations, pack sizes, or kit configurations may be available upon request.
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- Jakimiuk K, et al., 2021 J Enzyme Inhib Med Chem 36(1): 1016-1028.
