| << BACK TO SCREENING SERVICES LIST |
Inflammatory Screening – Migration Assay
The leukocyte adhesion cascade is a sequence of adhesion and activation events that ends with extravasation of the leukocyte, whereby the cell exerts its effects on the site of inflammation. The five steps of the adhesion cascade are capture, rolling, slow rolling, firm adhesion, and transmigration. Each of these five steps appears to be necessary for effective leukocyte recruitment and therefore constitute a target for anti-inflammatory drug discovery, whereby the goal is to develop methods to control inflammation by modulating or blocking leukocyte recruitment.
Assay Principles
GlycoMar’s inflammatory cell transmigration assay determines the migration of labelled human neutrophils through a monolayer of cultured Human umbilical vein endothelial cells (HUVECs). HUVECs are cultured to confluence on the filter of a transwell before being stimulated with IL-1ß and/or TNF-α. Test compounds are then added along with labeled leukocytes and the plate incubated for a set time to allow the cells to migrate. A chemotactic agent can also be added to the lower chamber at this point if required. Following incubation fluorescence readings are taken for the non-adherent, adherent (to the HUVEC layer) and migrated leukocytes.
Results
The percentage of leukocytes, relative to total leukocytes, is calculated for each of the three compartments – non-adherent (non-migrated), adherent, and migrated cells. Results are reported in a format and timescale agreed with the client. The assay is suitable for the performance of doseresponse studies.
GlycoMar is committed to ensuring quality in all of its activities through a GLP-based Quality Assurance system.
Figure 1.Transmigration Assay; % Leukocytes ± SEM. The graph shows stimulation of the monlayer results in increased adherence and migration of the neutrophils.
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 24-well transwell format offering low – medium throughput.
Standard turnaround is 4 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
| << BACK TO SCREENING SERVICES LIST |