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Anti-Oxidant Screening – Oxidative burst Assay
The oxidative burst is the rapid production of reactive oxygen species (ROS) and reactive nitrogen species (RNS) in a defensive inflammatory response, which plays an important role in the immune system of many organisms, including humans. Both ROS and RNS are widely believed to contribute to the development of several age-related diseases, such as cancer, atherosclerosis, and neurodegenerative disease, by causing oxidative stress or damage. The oxidative burst is an important process in inflammatory cell activation which offers a target for anti-inflammatory Research & Development.
Assay Principles
GlycoMar’s assay measures the production of reactive species (ROS and RNS) in freshly isolated human neutrophils, which are activated with the phorbol ester PMA. The amount of ROS / RNS produced is determined using a cell permeant fluorescent probe. This assay can be used to detect both anti-inflammatory and pro-inflammatory activity of test agents. In anti-inflammatory mode cells are stimulated with PMA and the assay measures the ability of test agents to inhibit production of reactive species. In pro-inflammatory mode cells are treated with the test agent only and the assay measures the ability of test agents to induce production of reactive species.
Results
ROS and RNS production is calculated as a percentage relative to a control cells. Results are reported, summarized graphically, in a format and timescale agreed with the client. The assay is suitable for the performance of dose-response studies.
Figure 1. Time course of reactive species production measured in human neutrophils, activated with PMA in the presence and absence of catalase (positive control). Mean fluorescence ± SEM.
Figure 2. Typical assay data showing % activity of activated neutrophils treated with test agents. Percentage activity is relative to PMA-stimulated control neutrophils ± coefficient of variation (CV).
GlycoMar is committed to ensuring quality in all of its activities through a GLP-based Quality Assurance system.
Sample Submission
Samples can be submitted to GlycoMar by prior arrangement, in a number of formats.
Pricing is dependent 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 3 weeks from receipt, but faster turnaround is possible on request.
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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