Description(s):
E-selectin is a transmembrane cell adhesion molecule that is expressed on endothelial cells in response to cytokines like IL-1β and TNFα. During inflammation, this protein plays an important role in recruiting leukocytes to the site of injury. E-selectin has been shown to bind to cancer cells of multiple tumor types, and it is believed to be involved in facilitating cancer metastasis.
Principle
The wells of the plate are coated with capture antibody specific to E-selectin. In this assay, the test sample initially reacts with the solid phase capture antibody, resulting in E-selectin being bound to the well. The wells are then washed to remove unbound proteins, and biotin-linked antibodies are added to bind to the immobilized E-selectin. After washing away the unbound antibodies, Streptavidin-HRP conjugate is added to form a complex with the antibody-bound E-selectin. After incubation, the wells are washed to remove unbound Streptavidin-HRP conjugate. TMB substrate is then added and forms a blue color when the HRP-linked antibodies are detected. The reaction is then terminated with Stop Solution, which changes the color from blue to yellow. The E-selectin concentration in each well is directly proportional to its color intensity and can be quantified by measuring its optical density at 450 nm (OD450) in a microplate reader.
