This assay measures membrane-bound E-Selectin expressed on stimulated HUVEC using enzyme-linked immunosorbent assay (ELISA). After stimulation of HUVEC (with E.coli LPS, for example), the cells are fixed and incubated with a specific mouse monoclonal primary antibody to human membrane-bound E- Selectin. The cells are then incubated with an anti-mouse IgG antibody conjugated with peroxidase to detect the primary Ab. A chromogen substrate solution is added to the wells and color develops in proportion to the amount of membrane-bound E-Selectin present on the cells. As the E-Selectin is membrane-bound, a protein concentration standard cannot be performed. Rather, this assay measures test sample stimulation HUVEC E-Selectin expression relative to positive and negative controls.
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