Description(s):
Galectin-3 is a member of the lectin family and contains a carbohydrate-recognition-binding domain that binds to β-galactoside. Due to its ability to bind to β-galactoside, galectin-3 plays an important role in cell-cell adhesion, cell-matrix interactions, macrophage activation, angiogenesis, metastasis, apoptosis. Given galectin-3's broad biological functionality, it has been demonstrated to be involved in a number of diseases including cancer, fibrosis, heart disease, and stroke. Overexpression of galectin-3 is commonly seen in cancerous conditions and can affect cancer cell growth and differentiation, chemoattraction, apoptosis, and metastasis. Because of this, galectin-3 is increasingly being used as a diagnostic marker for different cancers.
Principle
The wells of the plate are coated with capture antibody specific to galectin-3. In this assay, the test sample initially reacts with the solid phase capture antibody, resulting in galectin-3 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 galectin-3. After washing away the unbound antibodies, Streptavidin-HRP conjugate is added to form a complex with the antibody-bound galectin-3. 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 galectin-3 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.
