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Inflammatory Screening – Elastase Inhibition Assay
Neutrophil influx plays an important role in normal defence mechanisms at the site of inflammation, as well as contributing to the pathogenesis of various disorders. Neutrophil elastase, a major serine proteinase in man, is predominantly present in the azurophilic granules of neutrophils. Elastase is released at sites of inflammation and can cause severe tissue damage.
Assay Principles
GlycoMar’s Elastase binding assay is a cell-free assay which measures the inhibition of human neutrophil elastase (hNE) by test agents. Enzyme kinetics are measured using a colorimetric elastase-specific substrate in the presence of test agent and controls. For inhibition studies kinetic measurements are made over a range of substrate concentrations. For dose-response studies kinetic measurements are made over a range of test agent doses.
Results
For inhibition studies, kinetic data is used to generate Michaelis-Menten and Lineweaver-Burke plots, which provide characterisation of kinetic parameters, the identification of enzyme inhibitors, the characterisation of the mode of inhibition (competitive, non-competitive, etc.) and calculation of statistical significance.
Dose-response data is plotted to allow determination of IC50 values.
GlycoMar is committed to ensuring quality in all of its activities through a GLP-based Quality Assurance system.
Figure 1. Lineweaver-Burke plot of elastase inhibition by low molecular weight heparin.
Sample Submission
Samples can be submitted to GlycoMar by prior arrangement, in a number of formats.
Pricing is dependant on sample numbers submitted and turnaround required. Currently the assay is available in a 96-well microplate format offering low – medium throughput.
Standard turnaround is 2 weeks from receipt.
Please contact GlycoMar directly for a quotation, or to discuss your individual requirements:
Tel: +44 (0) 1631 559 368 E-mail: info@glycomar.com
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