Antigen-Down Assay Diluent is formulated for use with serum and plasma samples tested in antigen-down ELISA formats. Proprietary additives have been included to reduce the level of nonspecific IgG adsorption to antigen-coated and blocked ELISA well surfaces, as well as enhancing the specific anti-analyte/antigen-antibody signal without denaturing the plate adsorbed antigen molecules.
- Once the capture antigen has been coated onto the plate and the plate has been properly blocked with one of ICT’s stabilizing blocker products, pipette 50 – 100 µL of Antigen-Down Assay Diluent into each of the wells of the ELISA plate using a multi-channel pipettor. The Assay Diluent is always added prior to adding the test and control serum/plasma samples to plate wells.
- The control and test serum/plasma samples should be diluted to the proper well concentrations using one of ICT’s three different Sample Diluent products. Using an ICT Sample Diluent to dilute the serum and plasma samples will also serve to help minimize the degree of nonspecific IgG binding to the antigen coated and blocked ELISA plate wells. The test (sera and plasma) samples, diluted typically to 1:40 or greater in a sample diluent, should be incubated in the antigen coated ELISA plates for 30 – 60 minutes at room temperature or 37°C.