Leukocyte or tumor cell interactions with vascular endothelium consist of a cascade of processes including the firm attachment of cells to endothelial cell adhesion molecules.
The CytoSelect™ Leukocyte Endothelium Adhesion Assay provides a robust system for the quantitative determination of interactions between leukocytes and the endothelium. Adherent cells can be easily quantified on a fluorescence plate reader.
Assay Principle. Endothelial cells are cultured and form a monolayer, which is treated with desired activator or inhibitor. Meanwhile, leukocytes are labeled with the LeukoTracker™ solution, pelleted, washed and resuspended. Leukocytes are added to the monolayer and incubated. Adherent leukocytes are washed, lysed and quantitated by a fluorometer
Human Monocytic THP-1 Adhesion to HUVEC Monolayer. HUVEC monolayer in a 48-well plate was treated with 1 µM PMA for 12 hours. LeukoTracker™ labeled THP-1 cells were allowed to attach to HUVEC monolayer for 1 hour. Adherent cells were lysed and quantified as described in the assay protocol.